Note: Descriptions are shown in the official language in which they were submitted.
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ISOLATED HUMAN TRANSPORTER PROTEINS, NUCLEIC ACID MOLECULES
ENCODING HUMAN TRANSPORTER PROTEINS, AND USES THEREOF
FIELD OF THE INVENTION
The present invention is in the field of transporter proteins that are related
to the
synaptic vesicle protein subfamily, recombinant DNA molecules, and protein
production.
The present invention specifically provides novel peptides and proteins that
effect ligand
transport and nucleic acid molecules encoding such peptide and protein
molecules, all of
which are useful in the development of human therapeutics and diagnostic
compositions and
methods.
BACKGROUND OF THE INVENTION
Transporters
Transporter proteins regulate many different functions of a cell, including
cell
proliferation, differentiation, and signaling processes, by regulating the
flow of molecules
such as ions and macromolecules, into and out of cells. Transporters are found
in the plasma
membranes of virtually every cell in eukaryotic organisms. Transporters
mediate a variety
of cellular functions including regulation of membrane potentials and
absorption and
secretion of molecules and ion across cell membranes. When present in
intracellular
membranes of the Golgi apparatus and endocytic vesicles, transporters, such as
chloride
channels, also regulate organelle pH. For a review, see Greger, R. (1988)
Annu. Rev.
Physiol. 5 0:111-122.
Transporters are generally classified by structure and the type of mode of
action. In
addition, transporters are sometimes classified by the molecule type that is
transported, for
example, sugar transporters, chlorine channels, potassium channels, etc. There
may be many
classes of channels for transporting a single type of molecule (a detailed
review of channel
types can be found at Alexander, S.P.H. and J.A. Peters: Receptor and
transporter
nomenclature supplement. Trends Pharmacol. Sci., Elsevier, pp. 65-68 (1997)
and
http://www-biology.ucsd.edu/~msaier/transport/titlepa~e2.html.
The following general classification scheme is known in the art and is
followed in the
present discoveries.
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Channel-type transporters. Transmembrane chaiuiel proteins of this class are
ubiquitously found in the membranes of all types of organisms from bacteria to
higher ,,
eukaryotes. Transport systems of this type catalyze facilitated diffusion (by
an energy-
independent process) by passage through a transmembrane aqueous pore or
channel without
evidence for a carrier-mediated mechanism. These channel proteins usually
consist largely of
a-helical spanners, although b-strands may also be present and may even
comprise the
channel. However, outer membrane porin-type channel proteins are excluded from
this class
and are instead included in class 9.
Carrier-type transporters. Transport systems are included in this class if
they utilize a
carrier-mediated process to catalyze uniport (a single species is transported
by facilitated
diffusion), antiport (two or more species are transported in opposite
directions in a tightly
coupled process, not coupled to a direct form of energy other than
chemiosmotic energy) .
andlor symport (two or more species are transported together in the same
direction in a
tightly coupled process, not coupled to a direct form of energy other than
chemiosmotic
energy).
Pyrophosphate bond hydrolysis-driven active transporters. Transport systems
are
included in this class if they hydrolyze pyrophosphate or the terminal
pyrophosphate bond in
ATP or another nucleoside triphosphate to drive the active uptake and/or
extrusion of a
solute or solutes.' The transport protein may or may not be transiently
phosphorylated, but the
substrate is not phosphorylated.
PEP-dependent, phosphoryl transfer-driven group translocators. Transport
systems of
the bacterial phosphoenolpyruvateaugar phosphotransferase system are included
in this
class. The product of the reaction, derived from extracellular sugar, is a
cytoplasmic sugar-
phosphate.
Decarboxylation-driven active transporters. Transport systems that drive
solute (e.g.,
ion) uptake or extrusion by decarboxylation of a cytoplasmic substrate are
included in this
class.
Oxidoreduction-driven active transporters. Transport systems that drive
transport of a
solute (e.g., an ion) energized by the flow of electrons from a reduced
substrate to an
oxidized substrate are included in this class.
Light-driven active transporters. Transport systems that utilize light energy
to drive
transport of a solute (e.g., an ion) are included in this class.
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Mechanically-driven active transporters. Transport systems are included in
this class
if they drive movement of a cell or organelle by allowing the flow of ions (or
other solutes)
through the membrane down their electrochemical gradients.
Outer-membrane porins (of b-structure). These proteins form transmembrane
pores
or channels that usually allow the energy independent passage of solutes
across a membrane..
The transmembrane portions of these proteins consist exclusively of b-strands
that form a b-
barrel. These porin-type proteins are found in the outer membranes of Gram-
negative
bacteria, mitochondria and eukaryotic plastids.
Methyltransferase-driven active transporters. A single characterized protein
currently
falls into this category, the Na+-transporting
methyltetrahydromethanopterin:coenzyme M
methyltransferase.
Non-ribosome-synthesized channel-forming peptides or peptide-like molecules.
These molecules, usually chains of L- and D-amino acids as well as other small
molecular
building blocks such as lactate, form oligomeric transmembrane ion channels.
Voltage may
induce channel formation by promoting assembly of the transmembrane channel.
These
peptides are often made by bacteria and fungi as agents of biological warfare.
Non-Proteinaceous Transport Complexes. Ion conducting substances in biological
membranes that do not consist of or are not derived from proteins or peptides
fall into this
category.
Functionally characterized transporters for which sequence data are lacking.
Transporters of particular physiological significance will be included in this
category even
though a family assignment cannot be made.
Putative transporters in which no family member is an established transporter.
Putative transport protein families are grouped under this number and will
either be
classified elsewhere when the transport function of a member becomes
established, or will
be eliminated from the TC classification system if the proposed transport
function is
disproven. These families include a member or members for which a transport
function has
been suggested, but evidence for such a function is not yet compelling.
Auxiliary transport proteins. Proteins that in some way facilitate transport
across one
or more biological membranes but do not themselves participate directly in
transport are
included in this class. These proteins always function in conjunction with one
or more
transport proteins. They may provide a function connected with energy coupling
to transport,
play a structural role in complex formation or serve a regulatory function.
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Transporters of unknown classification. Transport protein families of unknown
classification are grouped under this number and will be classified elsewhere
when the
transport process and energy coupling mechanism are characterized. These
families include
at least one member for which a transport function has been established, but
either the mode
of transport or the energy coupling mechanism is not known.
Ion channels
An important type of transporter is the ion channel. Ion channels regulate
many
different cell proliferation, differentiation, and signaling processes by
regulating the flow of
ions into and out of cells. Ion channels are found in the plasma membranes of
virtually
every cell in eukaryotic organisms. Ion channels mediate a variety of cellular
functions
including regulation of membrane potentials and absorption and secretion of
ion across
epithelial membranes. When present in intracellular membranes of the Golgi
apparatus and
endocytic vesicles, ion channels, such as chloride channels, also regulate
organelle pH. Fox a
review, see Greger, R. (1988) Annu. Rev. Physiol. 50:111-122.
Ion channels are generally classified by structure and the type of mode of
action. For
example, extracellular ligand gated channels (ELGs) are comprised of five
polypeptide
subunits, with each subunit having 4 membrane spanning domains, and are
activated by the
binding of an extracellular ligand to the channel. In addition, channels are
sometimes
classified by the ion type that is transported, for example, chlorine
channels, potassium
channels, etc. There may be many classes of channels for transporting a single
type of ion (a
detailed review of channel types can be found at Alexander, S.P.H. and 1.A.
Peters (1997).
Receptor and ion channel nomenclature supplement. Trends Pharmacol. Sci.,
Elsevier, pp.
65-68 and http://www-biology.ucsd.edu/~msaier/transport/toc.html.
There are many types of ion channels based on structure. For example, many ion
channels fall within one of the following groups: extracellular ligand-gated
channels (ELG),
intracellular ligand-gated channels (ILG), inward rectifying channels (INR),
intercellular
(gap junction) channels, and voltage gated channels (VIC). There are
additionally
recognized other channel families based on ion-type transported, cellular
location and drug
sensitivity. Detailed information on each of these, their activity, ligand
type, ion type,
disease association, drugability, and other information pertinent to the
present invention, is
well known in the art.
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Extracellular ligand-gated channels, ELGs, are generally comprised of five
polypeptide subunits, Unwin, N. (1993), Cell 72: 31-41; Unwin, N. (1995),
Nature 373: 37-
43; Hucho, F., et al., (1996) J. Neurochem. 66: 1781-1792; Hucho, F., et al.,
(1996) Eur. J.
Biochem. 239: 539-557; Alexander, S.P.H. and J.A. Peters (1997), Trends
Pharmacol. Sci.,
Elsevier, pp. 4-6; 36-40; 42-44; and Xue, H. (1998) J. Mol. Evol. 47: 323-333.
Each subunit
has 4 membrane spanning regions: this serves as a means of identifying other
members of
the ELG family of proteins. ELG bind a ligand and in response modulate the
flow of ions.
Examples of ELG include most members of the neurotransmitter-receptor family
of proteins,
e.g., GABAI receptors. Other members of this family of ion channels include
glycine
receptors, ryandyne receptors, and ligand gated calcium channels.
The Voltage-dated Ion Channel (VIC) Superfamily
Proteins of the VIC family are ion-selective channel proteins found in a wide
range
of bacteria, archaea and eukaryotes Hille, B. (1992), Chapter 9: Structure of
channel
proteins; Chapter 20: Evolution and diversity. In: Ionic Channels of Excitable
Membranes,
2nd Ed., Sinaur Assoc. Inc., Pubs., Sunderland, Massachusetts; Sigworth, F.J.
(1993), Quart.
Rev. Biophys. 27: 1-40; Salkoff, L. and T. Jegla (1995), Neuron 15: 489-492;
Alexander,
S.P.H. et al., (1997), Trends Pharmacol. Sci., Elsevier, pp. 76-84; Jan, L.Y.
et al., (1997),
Annu. Rev. Neurosci. 20: 91-123; Doyle, D.A, et al., (1998) Science 280: 69-
77; Terlau, H.
and W. Stiihmer (1998), Naturwissenschaften 85: 437-444. They are often homo-
or
heterooligomeric structures with several dissimilar subunits (e.g., al-a2-d-b
Ca2+ channels,
ablb2 Na+ channels or (a)4-b K+ channels), but the channel and the primary
receptor is
usually associated with the a (or al) subunit. Functionally characterized
members are
specific for K+, Na or Ca2+. The K+ channels usually consist of homotetrameric
structures
with each a-subunit possessing six transmembrane spanners (TMSs). The al and a
subunits
of the Ca2+ and Nab channels, respectively, are about four times as large and
possess 4 units,
each with 6 TMSs separated by a hydrophilic loop, for a total of 24 TMSs.
These large
channel proteins form heterotetra-unit structures equivalent to the
homotetrameric structures
of most K+ channels. All four units of the Ca2+ and Na+ channels are
homologous to the
single unit in the homotetrameric K+ channels. Ion flux via the eukaryotic
channels is
generally controlled by the transmembrane electrical potential (hence the
designation,
voltage-sensitive) although some are controlled by ligand or receptor binding.
Several putative K+-selective channel proteins of the VIC family have been
identified
in prokaryotes. The structure of one of them, the KcsA K+ channel of
Streptomyces lividahs,
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has been solved to 3.2 ~ resolution. The protein possesses four identical
subunits, each with
two transmembrane helices, arranged in the shape of an inverted teepee or
cone. The cone
cradles the "selectivity filter" P domain in its outer end. The narrow
selectivity filter is only
12 ~ long, whereas the remainder of the channel is wider and lined with
hydrophobic
residues. A large water-filled cavity and helix dipoles stabilize K+ in the
pore. The selectivity
filter has two bound K+ ions about 7.5 A apart from each other. Ion conduction
is proposed
to result from a balance of electrostatic attractive and repulsive forces.
In eukaryotes, each VIC family channel type has several subtypes based on
pharmacological and elactrophysiological data. Thus, there are five types of
Caa+ channels
(L, N, P, Q and T). There are at least ten types of K+ channels, each
responding in different
ways to different stimuli: voltage-sensitive [Ka, Kv, Kvr, Kvs and Ksr], Ca2~-
sensitive
[BKCa, IKca and SKoa] and receptor-coupled [KM and KACn] ~ There are at least
six types of
Na channels (I, II, III, ~,1, H1 and PN3). Tetrameric channels from both
prokaryotic and
eukaryotic organisms are known in which each a-subunit possesses 2 TMSs rather
than 6,
and these two TMSs are homologous to TMSs 5 and 6 of the six TMS unit found in
the
voltage-sensitive channel proteins. KcsA of S. lividahs is an example of such
a 2 TMS
channel protein. These channels may include the KNa (Nab-activated) and Kvol
(cell volume-
sensitive) K+ channels, as well as distantly related channels such as the Tokl
K+ channel of
yeast, the TWIK-1 inward rectifier K+ channel of the mouse and the TREK-1 K+
channel of
the mouse. Because of insufficient sequence similarity with proteins of the
VIC family,
inward rectifier K+ IRK channels (ATP-regulated; G-protein-activated) which
possess a P
domain and two flanking TMSs are placed in a distinct family. However,
substantial
sequence similarity in the P region suggests that they are homologous. The b,
g and d
subunits of VIC family members, when present, frequently play regulatory roles
in channel
activation/deactivation.
The Epithelial Na Channel (ENaC) Family
The ENaC family consists of over twenty-four sequenced proteins (Canessa,
C.M., et
al., (1994), Nature 367: 463-467, Le, T. and M.H. Saier, Jr. (1996), Mol.
Membr. Biol. 13:
149-157; Garty, H. and L.G. Palmer (1997), Physiol. Rev. 77: 359-396;
Waldmann, R., et
al., (1997), Nature 386: 173-177; Darboux, L, et al., (1998), J. Biol. Chem.
273: 9424-9429;
Firsov, D., et al., (1998), EMBO J. 17: 344-352; Horisberger, J.-D. (1998).
Curr. Opin.
Struc. Biol. 10: 443-449). All are from animals with no recognizable
homologues in other
eukaryotes or bacteria. The vertebrate ENaC proteins from epithelial cells
cluster tightly
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together on the phylogenetic tree: voltage-insensitive ENaC homologues are
also found in
the brain. Eleven sequenced C. elegans proteins, including the degenerins, are
distantly
related to the vertebrate proteins as well as to each other. At least some of
these proteins
form part of a mechano-transducing complex for touch sensitivity. The
homologous Helix
aspersa (FMRF-amide)-activated Na+ channel is the first peptide
neurotransmitter-gated
ionotropic receptor to be sequenced.
Protein members of this family all exhibit the same apparent topology, each
with N-
and C-termini on the inside of the cell, two amphipathic transmembrane
spanning segments,
and a large extracellular loop. The extracellular domains contain numerous
highly conserved
cysteine residues. They are proposed to serve a receptor function.
Mammalian ENaC is important for the maintenance of Nab balance and the
regulation of blood pressure. Three homologous ENaC subunits, alpha, beta, and
gamma,
have been shown to assemble to form the highly Na +-selective channel. The
stoichiometry
of the three subunits is alpha2, betal, gammal in a heterotetrameric
architecture.
The Glutamate-gated Ion Channel (GIC) Family of Neurotransmitter Receptors
Members of the GIC family are heteropentameric complexes in which each of the
5
subunits is of 800-1000 amino acyl residues in length (Nakanishi, N., et al,
(1990), Neuron
5: 569-581; Unwin, N. (1993), Cell 72: 31-41; Alexander, S.P.H. and J.A.
Peters (1997)
Trends Pharmacol. Sci., Elsevier, pp. 36-40). These subunits may span the
membrane three
or five times as putative a-helices with the N-termini (the glutamate-binding
domains)
localized extracellulaxly and the C-termini localized cytoplasmically. They
may be distantly
related to the ligand-gated ion channels, and if so, they may possess
substantial b-structure in
their transmembrane regions. However, homology between these two families
cannot be
established on the basis of sequence comparisons alone. The subunits fall into
six
subfamilies: a, b, g, d, a and z.
The GIC channels are divided into three types: (1) a-amino-3-hydroxy-5-methyl-
4-
isoxazole propionate (AMPA)-, (2) kainate- and (3) N-methyl-D-aspartate (NMDA)-
selective glutamate receptors. Subunits of the AMPA and kainate classes
exhibit 35-40%
identity with each other while subunits of the NMDA receptors exhibit 22-24%
identity with
the former subunits. They possess large N-terminal, extracellulax glutamate-
binding domains
that are homologous to the periplasmic glutamine and glutamate receptors of
ABC-type
uptake permeases of Gram-negative bacteria. All known members of the GIC
family are
from animals. The different channel (receptor) types exhibit distinct ion
selectivities and
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conductance properties. The NMDA-selective large conductance channels are
highly
permeable to monovalent cations and Ca2+. The AMPA- and kainate-selective ion
channels
are permeable primarily to monovalent cations with only low permeability to
Ca2+.
The Chloride Channel (C1C, Family
The C1C family is a large family consisting of dozens of sequenced proteins
derived
from Gram-negative and Gram-positive bacteria, cyanobacteria, archaea, yeast,
plants and
animals (Steinmeyer, K., et al., (1991), Nature 354: 301-304; LTchida, S., et
al., (1993), J.
Biol. Chem. 268: 3821-3824; Huang, M.-E., et al., (1994), J. Mol. Biol. 242:
595-598;
Kawasaki, M., et al, (1994), Neuron 12: 597-604; Fisher, W.E., et al., (1995),
Genomics.
29:598-606; and Foskett, J.K. (1998), Annu. Rev. Physiol. 60: 689-717). These
proteins are
essentially ubiquitous, although they are not encoded within genomes of
Haemophilus
iufluenzae, Mycoplasma genitalium, and Mycoplasma pv~eumoniae. Sequenced
proteins vary
in size from 395 amino acyl residues (M. jan~zaschii) to 988 residues (man).
Several
organisms contain multiple C1C family paralogues. For example, Synechocystis
has two
paralogues, one of 451 residues in length and the other of 899 residues.
A~abidopsis thalia~a
has at least four sequenced paralogues, (775-792 residues), humans also have
at least five
paralogues (820-988 residues, and C. elegans also has at least five (810-950
residues).
There are nine known members in mammals, and mutations in three of the
corresponding
genes cause human diseases. E. coli, Metha~zococcus jahnaschii and
Saccharomyces
ce~evisiae only have one C1C family member each. With the exception of the
larger
Synechocystis paralogue, all bacterial proteins are small (395-492 residues)
while all
eukaryotic proteins are larger (687-988 residues). These proteins exhibit 10-
12 putative
transmembrane a-helical spanners (TMSs) and appear to be present in the
membrane as
homodimers. While one member of the family, Torpedo C1C-O, has been reported
to have
two channels, one per subunit, others are believed to have just one.
All functionally characterized members of the C1C family transport chloride,
some in
a voltage-regulated process. These channels serve a variety of physiological
functions (cell
volume regulation; membrane potential stabilization; signal transduction;
transepithelial
transport, etc.). Different homologues in humans exhibit differing anion
selectivities, i.e.,
C1C4 and C1C5 share a N03- > Cl- > Br > I- conductance sequence, while C1C3
has an I- >
Cl- selectivity. The C1C4 and C1C5 channels and others exhibit outward
rectifying currents
with currents only at voltages more positive than +20mV.
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Animal Inward Rectifier K~ Channel (IRK-C) Family
IRK channels possess the "minimal channel-forming structure" with only a P
domain,
characteristic of the channel proteins of the VIC family, and two flanking
transmembrane
spanners (Shuck, M.E., et al., (1994), J. Biol. Chem. 269: 24261-24270; Ashen,
M.D., et al.,
(1995), Am. J. Physiol. 268: H506-H511; Salkoff, L. and T. Jegla (I995),
Neuron 15: 489-
492; Aguilar-Bryan, L., et al., (1998), Physiol. Rev. 78: 227-245; Ruknudin,
A., et al.,
(1998), J. Biol. Chem. 273: 14165-14171). They may exist in the membrane as
homo- or
heterooligomers. They have a greater tendency to let K~ flow into the cell
than out. Voltage-
dependence may be regulated by external K+, by internal Mg2+, by internal ATP
and/or by
G-proteins. The P domains of IRK channels exhibit limited sequence similarity
to those of
the VIC family, but this sequence similarity is insufficient to establish
homology. Inward
rectifiers play a role in setting cellular membrane potentials, and the
closing of these
channels upon depolarization permits the occurrence of long duration action
potentials with a
plateau phase. Inward rectifiers lack the intrinsic voltage sensing helices
found in VIC
family channels. In a few cases, those of Kirl.la and Kir6.2, for example,
direct interaction
with a member of the ABC superfamily has been proposed to confer unique
functional and
regulatory properties to the heteromeric complex, including sensitivity to
ATP. The SURl
sulfonylurea receptor (spQ09428) is the ABC protein that regulates the Kir6.2
channel in
response to ATP, and CFTR may regulate Kirl.la. Mutations in SURl are the
cause of
familial persistent hyperinsulinemic hypoglycemia in infancy (PHHI), an
autosomal
recessive disorder characterized by unregulated insulin secretion in the
pancreas.
ATP-dated Cation Channel (ACC) Family
Members of the ACC family (also called P2X receptors) respond to ATP, a
functional neurotransmitter released by exocytosis from many types of neurons
(North; R.A.
(1996), Curr. Opin. Cell Biol. 8: 474-483; Soto, F., M. Garcia-Guzman and W.
Stiihmer
(1997), J. Membr. Biol. 160: 91-100). They have been placed into seven groups
(P2Xi -
P2X~) based on their pharmacological properties. These channels, which
function at neuron-
neuron and neuron-smooth muscle junctions, may play roles in the control of
blood pressure
and pain sensation. They may also function in lymphocyte and platelet
physiology. They are
found only in animals.
The proteins of the ACC family are quite similar in sequence (>35% identity),
but
they possess 380-1000 amino acyl residues per subunit with variability in
length localized
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primarily to the C-terminal domains. They possess two transmembrane spanners,
one about
30-50 residues from their N-termini, the other near residues 320-340. The
extracellular
receptor domains between these two spanners (of about 270 residues) are well
conserved
with numerous conserved glycyl and cysteyl residues. The hydrophilic C-termini
vary in
length from 25 to 240 residues. They resemble the topologically similar
epithelial Na
channel (ENaC) proteins in possessing (a) N- and C-termini localized
intracellularly, (b) two
putative transmembrane spanners, (c) a large extracellular loop domain, and
(d) many
conserved extracellular cysteyl residues. ACC family members are, however, not
demonstrably homologous with them. ACC channels are probably hetero- or
homomultimers
and transport small monovalent cations (Me~. Some also transport Ca2+; a few
also transport
small metabolites.
The Ryanodine-Inositol 1,4,5-tri_phosphate Receptor Ca2t Channel (RIR-CaC)
Familv
Ryanodine (Ry)-sensitive and inositol 1,4,5-triphosphate'(IP3)-sensitive Ca2~-
release
channels function in the release of Ca2+ from intracellular storage sites in
animal cells and
thereby regulate various Ca2+ -dependent physiological processes (Hasan, G. et
al., (1992)
Development 116: 967-975; Michikawa, T., et al., (1994), J. Biol. Chem. 269:
9184-9189;
Tunwell, R.E.A., (1996), Biochem. J. 318: 477-487; Lee, A.G. (1996)
Biomembrahes, Vol.
6, Transmembrane Receptors and Channels (A.G. Lee, ed.), JAI Press, Denver,
CO., pp 291-
326; Mikoshiba, K., et al., (1996) J. Biochem. Biomem. 6: 273-289). Ry
receptors occur
primarily in muscle cell sarcoplasmic reticular (SR) membranes, and IP3
receptors occur
primarily in brain cell endoplasmic reticular (ER) membranes where they effect
release of
Ca2+ into the cytoplasm upon activation (opening) of the channel.
The Ry receptors are activated as a result of the activity of dihydropyridine-
sensitive
Caa+ channels. The latter are members of the voltage-sensitive ion channel
(VIC) family.
Dihydropyridine-sensitive channels are present in the T-tubular systems of
muscle tissues.
Ry receptors are homotetrameric complexes with each subunit exhibiting a
molecular
size of over 500,000 daltons (about 5,000 amino acyl residues). They possess C-
terminal
domains with six putative transmembrane a -helical spanners (TMSs). Putative
pore-forming
sequences occur between the fifth and sixth TMSs as suggested for members of
the VIC
family. The large N-terminal hydrophilic domains and the small C-terminal
hydrophilic
domains are localized to the cytoplasm. Low resolution 3-dimensional
structural data are
available. Mammals possess at least three isoforms that probably arose by gene
duplication
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and divergence before divergence of the mammalian species. Homologues are
present in
humans and Caenorabditis elegans.
IP3 receptors resemble Ry receptors in many respects. (1) They are
homotetrameric
complexes with each subunit exhibiting a molecular size of over 300,000
daltons (about
2,700 amino acyl residues). (2) They possess C-terminal channel domains that
are
homologous to those of the Ry receptors. (3) The channel domains possess six
putative
TMSs and a putative channel lining region between TMSs 5 and 6. (4) Both the
large N-
terminal domains and the smaller C-terminal tails face the cytoplasm. (5) They
possess
covalently linked carbohydrate on extracytoplasmic loops of the channel
domains. (6) They
have three currently recognized isoforms (types 1, 2, and 3) in mammals which
are subject to
differential regulation and have different tissue distributions.
IP3 receptors possess three domains: N-terminal IP3-binding domains, central
coupling or regulatory domains and C-terminal channel domains. Channels are
activated by
IP3 binding, and like the Ry receptors, the activities of the IP3 receptor
channels are
regulated by phosphorylation of the regulatory domains, catalyzed by various
protein
kinases. They predominate in the endoplasmic reticular membranes of various
cell types in
the brain but have also been found in the plasma membranes of some nerve cells
derived
from a variety of tissues.
The channel domains of the Ry and IP3 receptors comprise a coherent family
that in
spite of apparent structural similarities, do not show appreciable sequence
similarity of the
proteins of the VIC family. The Ry receptors and the IP3 receptors cluster
separately on the
RIR-CaC family tree. They both have homologues in Drosophila. Based on the
phylogenetic
tree for the family, the family probably evolved in the following sequence:
(1) A gene
duplication event occurred that gave rise to Ry and IP3 receptors in
invertebrates. (2)
Vertebrates evolved from invertebrates. (3) The three isoforms of each
receptor arose as a
result of two distinct gene duplication events. (4) These isoforms were
transmitted to
mammals before divergence of the mammalian species.
The Organellar Chloride Channel (O-C1C~ FamilX
Proteins of the O-C1C family are voltage-sensitive chloride channels found in
intracellular membranes but not the plasma membranes of animal cells (Landry,
D, et al.,
(1993), J. Biol. Chem. 268: 14948-14955; Valenzuela, Set al., (1997), J. Biol.
Chem. 272:
12575-12582; and Duncan, R.R., et al., (1997), J. Biol. Chem. 272: 23880-
23886).
11
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They are found in human nuclear membranes, and the bovine protein targets to
the
microsomes, but not the plasma membrane, when expressed in Xev~opus laevis
oocytes.
These proteins are thought to function in the regulation of the membrane
potential and in
transepithelial ion absorption and secretion in the kidney. They possess two
putative
transmembrane a-helical spanners (TMSs) with cytoplasmic N- and C-termini and
a large
luminal loop that may be glycosylated. The bovine protein is 437 amino acyl
residues in
length and has the two putative TMSs at positions 223-239 and 367-385. The
human nuclear
protein is much smaller (241 residues). A C. elega~zs homologue is 260
residues long.
Synaptic Vesicle Proteins
The novel human protein, and encoding gene, provided by the present invention
is
related to the synaptic vesicle protein family of transporter proteins. The
protein of the
present invention shows the highest degree of similarity to rat synaptic
vesicle protein 2C
(SV2C), which is associated with synaptic vesicles and thought to be important
for transport
of neurotransmitters across membranes. The pxotein of the present invention is
also similar
to a synaptic vesicle protein known as SVOP (see Janz et al., J. Neurosci. 18
(22), 9269-
9281 (1998)). SV2C is highly glycosylated, whereas SVOP is not. SVOP is
expressed in
brain and endocxine cells where it is localized to synaptic vesicles and
microvesicles.
Furthermore, SVOP is expressed in all regions of the brain, with large
pyramidal neurons of
the cerebral cortex being the site of highest expression. It is thought that
SVOP is important
for uptake of a novel component of synaptic vesicles (Janz et al., J.
Neurosci. 18 (22), 9269-
9281 (1998)).
Due to their importance in neural physiology, particularly in regulating
transport at
synaptic vesicles, novel human synaptic vesicle proteins/genes, such as
provided by the
present invention, are valuable as potential targets for the development of
therapeutics to
treat neurological diseases/disorders, as well as other diseases/disorders.
Furthermore, SNPs
in synaptic vesicle protein genes, such as provided by the present invention,
may serve as
valuable markers for the diagnosis, prognosis, prevention, and/or treatment of
such
diseases/disorders.
Using the information provided by the present invention, reagents such as
probes/primers for detecting the SNPs or the expression of the protein/gene
provided herein
may be readily developed and, if desired, incorporated into kit formats such
as nucleic acid
12
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arrays, primer extension reactions coupled with mass spec detection (for SNP
detection), or
TaqMan PCR assays (Applied Biosystems, Foster City, CA).
Transporter proteins, particularly members of the synaptic vesicle protein
subfamily, are
a major target for drug action and development. Accordingly, it is valuable to
the field of
pharmaceutical development to identify and characterize previously unknown
transport
proteins. The present invention advances the state of the art by providing
previously
unidentified human transport proteins.
SUMMARY OF THE INVENTION
The present invention is based in part on the identification of amino acid
sequences
of human transporter peptides and proteins that are related to the synaptic
vesicle protein
subfamily, as well as allelic variants and other mammalian orthologs thereof.
These unique
peptide sequences, and nucleic acid sequences that encode these peptides, can
be used as
models for the development of human therapeutic targets, aid in the
identification of
therapeutic proteins, and serve as targets for the development of human
therapeutic agents
that modulate transporter activity in cells and tissues that express the
transporter.
Experimental data as provided in Figure 1 indicates expression in brain and
muscle.
DESCRIPTION OF THE FIGURE SHEETS
FIGURE 1 provides the nucleotide sequence of a cDNA molecule that encodes the
transporter protein of the present invention. (SEQ ID NO: l) In addition
structure and
functional information is provided, such as ATG start, stop and tissue
distribution, where
available, that allows one to readily determine specific uses of inventions
based on this
molecular sequence. Experimental data as provided iri Figure 1 indicates
expression in brain
and muscle.
FIGURE 2 provides the predicted amino acid sequence of the transporter of the
present invention. (SEQ ID N0:2) In addition structure and functional
information such as
protein family, function, and modification sites is provided where available,
allowing one to
readily determine specific uses of inventions based on this molecular
sequence.
FIGURE 3 provides genomic sequences that span the gene encoding the
transporter
protein of the present invention. (SEQ ID N0:3) In addition structure and
functional
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information, such as intron/exon structure, promoter location, etc., is
provided where
available, allowing one to readily determine specific uses of inventions based
on this
molecular sequence. As illustrated in Figure 3, SNPs were identified at 269
different
nucleotide positions.
DETAILED DESCRIPTION OF THE INVENTION
General Description
The present invention is based on the sequencing of the human genome. During
the
sequencing and assembly of the human genome, analysis of the sequence
information
revealed previously unidentified fragments of the human genome that encode
peptides that
share structural and/or sequence homology to protein/peptide/domains
identified and
characterized within the art as being a transporter protein or part of a
transporter protein and
are related to the synaptic vesicle protein subfamily. Utilizing these
sequences, additional
genomic sequences were assembled and transcript and/or cDNA sequences were
isolated and
characterized. Based on this analysis, the present invention provides amino
acid sequences
of human transporter peptides and proteins that are related to the synaptic
vesicle protein
subfamily, nucleic acid sequences in the form of transcript sequences, cDNA
sequences
andlor genomic sequences that encode these transporter peptides and proteins,
nucleic acid
variation (allelic; information), tissue distribution of expression, and
information about the
closest art known protein/peptide/domain that has structural or sequence
homology to the
transporter of the present invention.
In addition to being previously unknown, the peptides that are provided in the
present
invention are selected based on their ability to be used for the development
of commercially
important products and services. Specifically, the present peptides are
selected based on
homology and/or structural relatedness to known transporter proteins of the
synaptic vesicle
protein subfamily and the expression pattern observed. Experimental data as
provided in
Figure 1 indicates expression in brain and muscle. The art has clearly
established the
commercial importance of members of this family of proteins and proteins that
have
expression patterns similar to that of the present gene Some of the more
specific features of
the peptides of the present invention, and the uses thereof, are described
herein, particularly
in the Background of the Tnvention and in the annotation provided in the
Figures, and/or are
I4
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known within the art for each of the known synaptic vesicle protein family or
subfamily of
transporter proteins.
Specific Embodiments
Peptide Molecules
The present invention provides nucleic acid sequences that encode protein
molecules
that have been identif ed as being members of the transporter family of
proteins and are
related to the synaptic vesicle protein subfamily (protein sequences axe
provided in Figure 2,
transcript/cDNA sequences are provided in Figures 1 and genomic sequences are
provided in
Figure 3). The peptide sequences provided in Figure 2, as well as the obvious
variants
described herein, particularly allelic variants as identified herein and using
the information in
Figure 3, will be referred herein as the transporter peptides of the present
invention,
transporter peptides, or peptides/proteins of the present invention.
The present invention provides isolated peptide and protein molecules that
consist of,
consist essentially of, or comprising the amino acid sequences of the
transporter peptides
disclosed in the Figure 2, (encoded by the nucleic acid molecule shown in
Figure 1,
transcript/cDNA or Figure 3, genomic sequence), as well as all obvious
variants of these
peptides that are within the art to make and use. Some of these variants are
described in
detail below.
As used herein, a peptide is said to be "isolated" or "purified" when it is
substantially
free of cellular material or free of chemical precursors or other chemicals.
The peptides of the
present invention can be purified to homogeneity or other degrees of purity.
The level of
purification will be based on the intended use. The critical feature is that
the preparation allows
for the desired function of the peptide, even if in the presence of
considerable amounts of other
components (the features of an isolated nucleic acid molecule is discussed
below).
In some uses, "substantially free of cellular material" includes preparations
of the
peptide having less than about 30% (by dry weight) other proteins (i.e.,
contaminating protein),
less than about 20% other proteins, less than about 10% other proteins, or
less than about 5%
other proteins. When the peptide is recombinantly produced, it can also be
substantially free of
culture medium, i.e., culture medium represents less than about 20% of the
volume of the
protein preparation.
CA 02439401 2003-08-25
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The language "substantially free of chemical precursors or other chemicals"
includes
preparations of the peptide in which it is separated from chemical precursors
or other chemicals
that are involved in its synthesis. In one embodiment, the language
"substantially free of
chemical precursors or other chemicals" includes preparations of the
transporter peptide having
less than about 30% (by dry weight) chemical precursors or other chemicals,
less than about
20% chemical precursors or other chemicals, less than about 10% chemical
precursors or other
chemicals, or less than about 5% chemical precursors or other chemicals.
The isolated transporter peptide can be purified from cells that naturally
express it,
purified from cells that have been altered to express it (recombinant), or
synthesized using
known protein synthesis methods. Experimental data as provided in Figure 1
indicates
expression in brain and muscle. For example, a nucleic acid molecule encoding
the transporter
peptide is cloned into an expression vector, the expression vector introduced
into a host cell and
the protein expressed in the host cell. The protein can then be isolated from
the cells by an
appropriate purification scheme using standard protein purification
techniques. Many of these
techniques are described in detail below.
Accordingly, the present invention provides proteins that consist of the amino
acid
sequences provided in Figure 2 (SEQ ID N0:2), for example, proteins encoded by
the
transcript/cDNA nucleic acid sequences shown in Figure 1 (SEQ ID NO:1) and the
genomic
sequences provided in Figure 3 (SEQ 1D N0:3). The amino acid sequence of such
a protein is
provided in Figure 2. A protein consists of an amino acid sequence when the
amino acid
sequence is the final axnino acid sequence of the protein.
The present invention further provides proteins that consist essentially of
the amino acid
sequences provided in Figure 2 (SEQ ID N0:2), for example, proteins encoded by
the
transcript/cDNA nucleic acid sequences shown in Figure 1 (SEQ ID NO:l) and the
genomic
sequences provided in Figure 3 (SEQ ID N0:3). A protein consists essentially
of an amino acid
sequence when such an amino acid sequence is present with only a few
additional amino acid
residues, for example from about 1 to about 100 or so additional residues,
typically from 1 to
about 20 additional residues in the final protein.
The present invention further provides proteins that comprise the amino acid
sequences
provided in Figure 2 (SEQ ID N0:2), for example, proteins encoded by the
transcript/cDNA
nucleic acid sequences shown in Figure 1 (SEQ ID NO:1) and the genomic
sequences provided
in Figure 3 (SEQ ID N0:3). A protein comprises an amino acid sequence when the
amino acid
sequence is at least part of the final amino acid sequence of the protein. Tn
such a fashion, the
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CA 02439401 2003-08-25
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protein can be only the peptide or have additional amino acid molecules, such
as amino acid
residues (contiguous encoded sequence) that are naturally associated with it
or heterologous
amino acid residues/peptide sequences. Such a protein can have a few
additional amino acid
residues or can comprise several hundred or more additional amino acids. The
preferred classes
of proteins that are comprised of the transporter peptides of the present
invention are the ,
naturally occurring mature proteins. A brief description of how various types
of these proteins
can be made/isolated is provided below.
The transporter peptides of the present invention can be attached to
heterologous
. sequences to form chimeric or fusion proteins. Such chimeric and fusion
proteins comprise a
transporter peptide operatively linked to a heterologous protein having an
amino acid sequence
not substantially homologous to the transporter peptide. "Operatively linked"
indicates that the
transporter peptide and the heterologous protein are fused in-frame. The
heterologous protein
can be fused to the N-terminus or C-terminus of the transporter peptide.
In some uses, the fusion protein does not affect the activity of the
transporter peptide
per se. For example, the fusion protein can include, but is not limited to,
enzymatic fusion
proteins, for example beta-galactosidase fusions, yeast two-hybrid GAL
fusions, poly-His
fusions, MYC-tagged, HI-tagged and Ig fusions. Such fusion proteins,
particularly poly-His
fusions, can facilitate the purification of recombinant transporter peptide.
In certain host cells
(e.g., .mammalian host cells), expression and/or secretion of a protein can be
increased by using
a heterologous signal sequence.
A chimeric or fusion protein can be produced by standard recombinant DNA
techniques. For example, DNA fragments coding for the different protein
sequences are ligated
together in-frame in accordance with conventional techniques. In another
embodiment, the
fusion gene can be synthesized by conventional techniques including automated
DNA
synthesizers. Alternatively, PCR amplification of gene fragments can be
carried out using
anchor primers which give rise to complementary overhangs between two
consecutive gene
fragments which can subsequently be annealed and re-amplified to generate a
chimeric gene
sequence (see Ausubel et al., Cu~re~t Protocols i~t Molecular Biology, 1992).
Moreover, many
expression vectors are commercially available that already encode a fusion
moiety (e.g., a GST
protein). A transporter peptide-encoding nucleic acid can be cloned into such
an expression
vector such that the fusion moiety is linked in-frame to the transporter
peptide.
As mentioned above, the present invention also provides and enables obvious
variants
of the amino acid sequence of the proteins of the present invention, such as
naturally occurring
17
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WO 02/083900 PCT/US02/05520
mature forms of the peptide, allelic/sequence variants of the peptides, non-
naturally occurring
recombinantly derived variants of the peptides, and orthologs and paralogs of
the peptides.
Such variants can readily be generated using art-known techniques in the
fields of recombinant
nucleic acid technology and protein biochemistry. It is understood, however,
that variants
exclude any amino acid sequences disclosed prior to the invention.
Such variants can readily be identified/made using molecular techniques and
the
sequence information disclosed herein. Further, such variants can readily be
distinguished
from other peptides based on sequence and/or structural homology to the
transporter peptides of
the present invention. The degree of homology/identity present will be based
primarily on
whether the peptide is a functional variant or non-functional vaxiant, the
amount of divergence
present in the paralog family and the evolutionary distance between the
orthologs.
To determine the percent identity of two amino acid sequences or two nucleic
acid
sequences, the sequences are aligned for optimal comparison purposes (e.g.,
gaps can be ,
introduced in one or both of a first and a second amino acid or nucleic acid
sequence for
optimal alignment and non-homologous sequences can be disregarded for
comparison
purposes). In a preferred embodiment, at least 30%, 40%, 50%, 60%, 70%, 80%,
or 90% or
more of a reference sequence is aligned for comparison purposes. The amino
acid residues
or nucleotides at corresponding amino acid positions or nucleotide positions
are then
compared. When a position in the first sequence is occupied by the same amino
acid xesidue
or nucleotide as the corresponding position in the second sequence, then the
molecules are
identical at that position (as used herein amino acid or nucleic acid
"identity" is equivalent to
amino acid or nucleic acid "homology"). The percent identity between the two
sequences is
a function of the number of identical positions shared by the sequences,
taking into account
the number of gaps, and the length of each gap, which need to be introduced
for optimal
alignment of the two sequences.
The comparison of sequences and determination of percent identity and
similarity
between two sequences can be accomplished using a mathematical algorithm.
(Computational Molecular Biology, Lesk, A.M., ed., Oxford University Press,
New York,
1988; Biocomputing: Ihfo~matics aid Gehome Projects, Smith, D.W., ed.,
Academic Press,
New York, 1993; ComputerA~alysis ofSequence Data, Part 1, Griffin, A.M., and
Griffin,
H.G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular
Biology, von
Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M.
and Devereux,
J., eds., M Stockton Press, New York,1991). In a preferred embodiment, the
percent identity
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WO 02/083900 PCT/US02/05520
between two amino acid sequences is determined using the Needleman and Wunsch
(J. Mol.
Biol. (48):444-453 (1970)) algorithm which has been incorporated into the GAP
program in
the GCG software package (available at http://www.gcg.com), using either a
Blossom 62
matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and
a length
weight of 1, 2, 3, 4, 5, or 6. In yet another preferred embodiment, the
percent identity
between two nucleotide sequences is determined using the GAP program in the
GCG
software package (Devereux, J., et al., NucleicAcids Res. 12(1):387 (1984))
(available at
http://www.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50,
60, 70,
or 80 and a length weight of 1, 2, 3, 4~ 5, or 6. In another embodiment, the
percent identity
between two amino acid or nucleotide sequences is determined using the
algorithm of E.
Myers and W. Miller (CABIOS, 4:11-17 (I989~) which has been incorporated info
the
ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length
penalty
of 12 and a gap penalty of 4.
The nucleic acid and protein sequences of the present invention can further be
used
, as a "query sequence" to perform a search against sequence databases to, for
example,
identify other family members or related sequences. Such searches can be
performed using
the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (J. Mol.
Biol. 215:403-
10 (1990)). BLAST nucleotide searches can be performed with the NBLAST
program,
score = 100, wordlength = 12 to obtain nucleotide sequences homologous to the
nucleic acid
molecules of the invention. BLAST protein searches can be performed with the
XBLAST
program, score = 50, wordlength = 3 to obtain amino acid sequences homologous
to the
proteins of the invention. To obtain gapped alignments for comparison
purposes, Gapped
BLAST can be utilized as described in Altschul et. al. (Nucleic Acids Res.
25(17):3389-3402
(1997)). When utilizing BLAST and gapped BLAST programs, the default
parameters of
the respective programs (e.g., XBLAST and NBLAST) can be used.
Full-length pre-processed forms, as well as mature processed forms, of
proteins that
comprise one of the peptides of the present invention can readily be
identified as having
complete sequence identity to one of the transporter peptides of the present
invention as well as
being encoded by the same genetic locus as the transporter peptide provided
herein. The gene
encoding the novel transporter protein of the present invention is located on
a genome
component that has been mapped to human chromosome 5 (as indicated in Figure
3), which is
supported by multiple lines of evidence, such as STS and BAC map data.
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Allelic variants of a transporter peptide can readily be identified as being a
human
protein having a high degree (significant) of sequence homology/identity to at
least a portion of
the transporter peptide as well as being encoded by the same genetic locus as
the transporter
peptide provided herein. Genetic locus can readily be determined based on the
genomic
information provided in Figure 3, such as the genomic sequence mapped to the
reference
. human. The gene encoding the novel transporter protein of the present
invention is located on a
genome component that has been mapped to human chromosome 5 (as indicated in
Figure 3),
which is supported by multiple lines of evidence, such as STS and BAC map
data. As used
herein, two proteins (or a region of the proteins) have significant homology
when the amino
acid sequences are typically at least about 70-80%, 80-90%, and more typically
at least about
90-95% or more homologous. A significantly homologous amino acid sequence,
according
to the present invention, will be encoded by a nucleic acid sequence that will
hybridize to a
transporter peptide encoding nucleic acid molecule under stringent conditions
as more fully
described below. _
Figure 3 provides information on SNPs that have been found in the gene
encoding
the transporter protein of the present invention. SNPs were identified at 269
different
nucleotide positions, including non-synonymous coding SNPs at positions
166328, 169076,
171899, and 171919. Changes in the amino acid sequence caused by these SNPs is
indicated
in Figure 3 and can readily be determined using the universal genetic code and
the protein
sequence provided in Figure 2 as a reference. Some of the SNPs that axe
located outside the
ORF and in introns may affect gene transcription.
Paralogs of a transporter peptide can readily be identified as having some
degree of
significant sequence homology/identity to at least a portion of the
transporter peptide, as being
encoded by a gene from humans, and as having similar activity or function. Two
proteins will
typically be considered paralogs when the amino acid sequences are typically
at least about
60% or greater, and more typically at least about 70% or greater homology
through a given
region or domain. Such paralogs will be encoded by a nucleic acid sequence
that will
hybridize to a transporter peptide encoding nucleic acid molecule under
moderate to
stringent conditions as more fully described below.
Orthologs of a transporter peptide can readily be identified as having some
degree of
significant sequence homology/identity to at least a portion of the
transporter peptide as well as
being encoded by a gene from another organism. Preferred orthologs will be
isolated from
mammals, preferably primates, for the development of human therapeutic targets
and agents.
CA 02439401 2003-08-25
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Such orthologs will be encoded by a nucleic acid sequence that will hybridize
to a transporter
peptide encoding nucleic acid molecule under moderate to stringent conditions,
as more fully
described below, depending on the degree of relatedness of the two organisms
yielding the
proteins.
Non-naturally occurring variants of the transporter peptides of the present
invention can
readily be generated using recombinant techniques. Such variants include, but
are not limited
to deletions, additions and substitutions in the amino acid sequence of the
transporter peptide.
For example, one class of substitutions are conserved amino acid substitution.
Such
substitutions are those that substitute a given amino acid in a transporter
peptide by another
amino acid of like characteristics. Typically seen as conservative
substitutions are the
replacements, one for another, among the aliphatic amino acids Ala, Val, Leu,
and Ile;
interchange of the hydroxyl residues Ser and,Thr; exchange of the acidic
residues Asp and Glu;
substitution between the amide residues Asn and Gln; exchange of the basic
residues Lys and .
Arg; and replacements among the aromatic residues Phe and Tyr. Guidance
concerning which
amino acid changes are likely to be phenotypically silent are found in Bowie
et al., Science
247:1306-1310 (1990).
Variant transporter peptides can be fully functional or can lack function in
one or more
activities, e.g. ability to bind ligand, ability to transport ligand, ability
to mediate signaling, etc.
Fully functional variants typically contain only conservative variation or
variation in non-
critical residues or in non-critical regions. Figure 2 provides the result of
protein analysis and
can be used to identify critical domains/regions. Functional variants can also
contain
substitution of similar amino acids that result in no change or an
insignificant change in
function. Alternatively, such substitutions may positively or negatively
affect function to some
degree.
Non-functional variants typically contain one or more non-conservative amino
acid
substitutions, deletions, insertions, inversions, or truncation or a
substitution, insertion,
inversion, or deletion in a critical residue or critical region.
.Amino acids that are essential for function can be identified by methods
known in the
art, such as site-directed mutagenesis or alanine-scanning mutagenesis
(Cunningham et al.,
Science 244:1081-1085 (1989)), particularly using the results provided in
Figure 2. The latter
procedure introduces single alanine mutations at every residue in the
molecule. The resulting
mutant molecules are then tested for biological activity such as transporter
activity or in assays
such as an in vitro proliferative activity. Sites that are critical for
binding parfner/substrate
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binding can also be determined by structural analysis such as crystallization,
nuclear magnetic
resonance or photoaffinity labeling (Smith et al., J. Mol. Biol. 224:899-904
(1992); de Vos et
al. Science 255:306-312 (1992)).
The present invention further provides fragments of the transporter peptides,
in addition
to proteins and peptides that comprise and consist of such fragments,
particularly those
comprising the residues identified in Figure 2. The fragments to which the
invention pertains,
however, are not to be construed as encompassing fragments that may be
disclosed publicly
prior to the present invention.
As used herein, a fragment comprises at least 8, 10, 12, 14,16, or more
contiguous
amino acid residues from a transporter peptide. Such fragments can be chosen
based on the
ability to retain one or more of the biological activities of the transporter
peptide or could be
chosen for the ability to perform a function, e.g. bind a substrate or act as
an immunogen.
Particularly important fragments are biologically active fragments, peptides
that are, for
example, about 8 or more amino acids in length. Such fragments will typically
comprise a
domain or motif of the transporter peptide, e.g., active site, a transmembrane
domain or a
substrate-binding domain. Further, possible fragments include, but are not
limited to, domain
or motif containing fragments, soluble peptide fragments, and fragments
containing
immunogenic structures. Predicted domains and functional sites are readily
identifiable by
computer programs well known and readily available to those of skill in the
art (e.g., PROSITE
analysis). The results of one such analysis are provided in Figure 2.
Polypeptides often contain amino acids other than the 20 amino acids commonly
referred to as the 20 naturally occurring amino acids. Further, many amino
acids, including the
terminal amino acids, may be modified by natural processes, such as processing
and other post-
translational modifications, or by chemical modification techniques well known
in the art.
Common modifications that occur naturally in transporter peptides are
described in basic texts,
detailed monographs, and the research literature, and they are well known to
those of skill in the
art (some of these features are identified in Figure 2).
Known modifications include, but are not limited to, acetylation, acylation,
ADP-
ribosylation, amidation, covalent attachment of flavin, covalent attachment of
a heme moiety,
covalent attachment of a nucleotide or nucleotide derivative, covalent
attachment of a lipid or
lipid derivative, covalent attachment of phosphotidylinositol, cross-linking,
cyclization,
disulfide bond formation, demethylation, formation of covalent crosslinks,
formation of cystine,
formation of pyroglutamate, formylation, gamma carboxylation, glycosylation,
GPI anchor
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CA 02439401 2003-08-25
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formation, hydroxylation, iodination, methylation, myristoylation, oxidation,
proteolytic
processing, phosphorylation, prenylation, racemization, selenoylation,
sulfation, transfer-RNA
mediated addition of amino acids to proteins such as arginylation, and
ubiquitination.
Such modifications are well known to those of skill in the art and have been
described
in great detail in the scientific literature. Several particularly common
modifications,
glycosylation, lipid attachment, sulfation, gamma-carboxylation of glutamic
acid residues,
hydroxylation and ADP-ribosylation, for instance, are described in most basic
texts, such as
Proteins - Structure and Molecular Properties, 2nd Ed., T.E. Creighton, W. H.
Freeman and
Company, New York (1993). Many detailed reviews are available on this subject,
such as by
Wold, F., Posttranslational Covale~at Mod~catioh of Proteins, B.C. Johnson,
Ed., Academic
Press, New York 1-12 (1983); Seifter et al. (Meth. En~ymol. 182: 626-646
(1990)) and Rattan et
al. (Arcn. N. Y. Acad. Sci. 663:48-62 (1992)).
Accordingly, the transporter peptides of the present invention also encompass
derivatives or analogs in which a substituted amino acid residue is not one
encoded by the
genetic code, in which a substituent group is included, in which the mature
transporter peptide
is fused with another compound, such as a compound to increase the half life
of the transporter
peptide (for example, polyethylene glycol), or in which the additional amino
acids are fused to
the mature transporter peptide, such as a leader or secretory sequence or a
sequence for
purification of the mature transporter peptide or a pro-protein sequence.
Protein/Peptide Uses
The proteins of the present invention can be used in substantial and specific
assays
related to the functional information provided in the Figures; to raise
antibodies or to elicit
another immune response; as a reagent (including the labeled reagent) in
assays designed to
quantitatively determine levels of the protein (or its binding partner or
ligand) in biological
fluids; and as markers for tissues in which the corresponding protein is
preferentially
expressed (either constitutively or at a particular stage of tissue
differentiation or
development or in a disease state). Where the protein binds or potentially
binds to another
protein or ligand (such as, for example, in a transporter-effector protein
interaction or
transporter-ligand interaction), the protein can be used to identify the
binding partner/ligand
so as to develop a system to identify inhibitors of the binding interaction.
Any or all of these
uses are capable of being developed into reagent grade or kit format for
commercialization
as commercial products.
23
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Methods for performing the uses listed above are well known to those skilled
in the
art. References disclosing such methods include "Molecular Cloning: A
Laboratory
Manual", 2d ed., Cold Spring Harbor Laboratory Press, Sambrook, J., E. F.
Fritsch and T.
Maniatis eds., 1989, and "Methods in Enzymology: Guide to Molecular Cloning
Techniques", Academic Press, Berger, S. L. and A. R. Kimmel eds., 1987.
The potential uses of the peptides of the present invention are based
primarily on the
source of the protein as well as the class/action of the protein. For example,
transporters
isolated from humans and their human/mammalian orthologs serve as targets for
identifying
agents for use in mammalian therapeutic applications, e.g. a human drug,
particularly in
modulating a biological or pathological response in a cell or tissue that
expresses the
transporter. Experimental data as provided in Figure 1 indicates that the
transporter proteins
of the present invention are expressed in brain and muscle, as indicated by
virtual northern
blot analysis. A large percentage of pharmaceutical agents are being developed
that
modulate the activity of transporter proteins, particularly members of the
synaptic vesicle
protein subfamily (see Background of the Invention). The structural and
functional
information provided in the Background and Figures provide specific and
substantial uses
for the molecules of the present invention, particularly in combination with
the expression
information provided in Figure 1. Experimental data as provided in Figure 1
indicates
expression in brain and muscle. Such uses can readily be determined using the
information
provided herein, that known in the art and routine experimentation.
The proteins of the present invention (including variants and fragments that
may have
been disclosed prior to the present invention) are useful for biological
assays related to
transporters that are related to members of the synaptic vesicle protein
subfamily. Such assays
involve any of the known transporter functions or activities or properties
useful for diagnosis
and treatment of transporter-related conditions that are specific for the
subfamily of transporters
that the one of the present invention belongs to, particularly in cells and
tissues that express the
transporter. Experimental data as provided in Figure 1 indicates that the
transporter proteins
of the present invention are expressed in brain and muscle, as indicated by
virtual northern
blot analysis. The proteins of the present invention are also useful in drug
screening assays, in
cell-based or cell-free systems ((Hodgson, Biotechnology, 1992, Sept 10(9);973-
80). Cell-
based systems can be native, i.e., cells that normally express the
transporter, as a biopsy or
expanded in cell culture. Experimental data as provided in Figure 1 indicates
expression in
24
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brain and muscle. In an alternate embodiment, cell-based assays involve
recombinant host cells
expressing the transporter protein.
The polypeptides can be used to identify compounds that modulate transporter
activity
of the protein in its natural state or an altered form that causes a specific
disease or pathology
associated with the transporter. Both the transporters of the present
invention and appropriate
variants and fragments can be used in high-throughput screens to assay
candidate compounds
for the ability to bind to the transporter. These compounds can be further
screened against a
functional transporter to determine the effect of the compound on the
transporter activity.
Further, these compounds can be tested in animal or invertebrate systems to
determine
activity/efFectiveness. Compounds can be identified that activate (agonist) or
inactivate
(antagonist) the transporter to a desired degree.
Further, the proteins of the present invention can be used to screen a
compound for the
ability to stimulate or inhibit interaction between the transporter protein
and a molecule that
normally interacts with the transporter protein, e.g. a substrate or a
component of the signal
I S pathway that the transporter protein normally interacts (for example,
another transporter). Such
assays typically include the steps of combining the transporter protein with a
candidate
compound under conditions that allow the transporter protein, or fragment, to
interact with the
target molecule, and to detect the formation of a complex between the protein
and the target or
to detect the biochemical consequence of the interaction with the transporter
protein and the
target, such as any of the associated effects of signal transduction such as
changes in membrane
potential, protein phosphorylation, cAMP turnover, and adenylate cyclase
activation, etc.
Candidate compounds include, for example, 1) peptides such as soluble
peptides,
including Ig-tailed fusion peptides and members of random peptide libraries
(see, e.g., Lam et
al., Nature 354:82-84 (1991); Houghten et al., Nature 354:84-86 (1991)) and
combinatorial
chemistry-derived molecular libraries made of D- and/or L- configuration amino
acids; 2)
phosphopeptides (e.g., members of random and partially degenerate, directed
phosphopeptide
libraries, see, e.g., Songyang et al., Cell 72:767-778 (1993)); 3) antibodies
(e.g., polyclonal,
monoclonal, humanized; anti-idiotypic, chimeric, and single chain antibodies
as well as Fab,
F(ab')2, Fab expression library fragments, and epitope-binding fragments of
antibodies); and 4)
small organic and inorganic molecules (e.g., molecules obtained from
combinatorial and natural
product libraries).
One candidate compound is a soluble fragment of the receptor that competes for
ligand
binding. Other candidate compounds include mutant transporters or appropriate
fragments
CA 02439401 2003-08-25
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containing mutations that affect transporter fiulction and thus compete for
ligand. Accordingly,
a fragment that competes for ligand, for example with a higher affinity, or a
fragment that binds
ligand but does not allow release, is encompassed by the invention.
The invention further includes other end point assays to identify compounds
that
modulate (stimulate or inhibit) transporter activity. The assays typically
involve an assay of
events in the signal transduction pathway that indicate transporter activity.
Thus, the transport
of a ligand, change in cell membrane potential, activation of a protein, a
change in the
expression of genes that are up- or down-regulated in response to the
transporter protein
dependent signal cascade can be assayed.
Any of the biological or biochemical functions mediated by the transporter can
be used
as an endpoint assay. These include all of the biochemical or
biochemical/biological events
described herein, in the references cited herein, incorporated by reference
for these endpoint
assay targets, and other functions known to those of ordinary skill in the art
or that can be
readily identified using the information provided in the Figures, particularly
Figure 2. ,
~ Specifically, a biological function of a cell or tissues that expresses the
transporter can be
assayed. Experimental data as provided in Figure 1 indicates that the
transporter proteins of
the present invention are expressed in brain and muscle, as indicated by
virtual northern blot
analysis.
Binding and/or activating compounds can also be screened by using chimeric
~0 transporter proteins in which the amino terminal extracellular domain, or
parts thereof, the
entire transmembrane domain or subregions, such as any of the seven
transmembrane segments
or any of the intracellular or extracellular loops and the carboxy terminal
intracellular domain,
or parts thereof, can be replaced by heterologous domains or subregions. For
example; a
ligand-binding region can be used that interacts with a different ligand then
that which is
recognized by the native transporter. Accordingly, a different set of signal
transduction
components is available as an end-point assay for activation. This allows for
assays to be
performed in other than the specific host cell from which the transporter is
derived.
The proteins of the present invention are also useful in competition binding
assays in
methods designed to discover compounds that interact with the transporter
(e.g. binding
partners and/or ligands). Thus, a compound is exposed to a transporter
polypeptide under
conditions that allow the compound to bind or to otherwise interact with the
polypeptide.
Soluble transporter polypeptide is also added to the mixture. If the test
compound interacts with
the soluble transporter polypeptide, it decreases the amount of complex formed
or activity from
26
CA 02439401 2003-08-25
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the transporter target. This type of assay is particularly useful in cases in
which compounds are
sought that interact with specific regions of the transporter. Thus, the
soluble polypeptide that
competes with the target transporter region is designed to contain peptide
sequences
corresponding to the region of interest.
To perform cell free drug screening assays, it is sometimes .desirable to
immobilize
either the transporter protein, or fragment, or its target molecule to
facilitate separation of
complexes from uncomplexed forms of one or both of the proteins; as well as to
accommodate
automation of the assay.
Techniques for immobilizing proteins on matrices can be used in the drug
screening
assays. In one embodiment, a fusion protein can be provided which adds a
domain that allows
the protein to be bound to a matrix. For example, glutathione-S-transferase
fusion proteins can
be adsorbed onto glutathione sepharose beads (Sigma Chemical; St. Louis, MO)
or glutathione
derivatized microtitre plates, which are then combined with the cell lysates
(e.g., 35S-labeled)
and the candidate-compound, and the mixture incubated under conditions
conducive to complex
formation (e.g., at physiological conditions for salt and pI-~. Following
incubation, the beads
are washed to remove any unbound label, and the matrix immobilized and
radiolabel
determined directly, or in the supernatant after the complexes are
dissociated. Alternatively, the
complexes can be dissociated from the matrix, separated by SDS-PAGE, and the
level of
transporter-binding protein found in the bead fraction quantitated from the
gel using standard
electrophoretic techniques. For example, either the polypeptide or its target
molecule can be
immobilized~utilizing conjugation of biotin and streptavidin using techniques
well known in the
art. Alternatively, antibodies reactive with the protein but which do not
interfere with binding
of the protein to its target molecule can be derivatized to the wells of the
plate, and the protein
trapped in the wells by antibody conjugation. Preparations of a transporter-
binding protein and
a candidate compound are incubated in the transporter protein-presenting wells
and the amount
of complex trapped in the well can be quantitated. Methods for detecting such
complexes, in
addition to those described above for the GST-immobilized complexes, include
immunodetection of complexes using antibodies reactive with the transporter
protein target
molecule, or which are reactive with transporter protein and compete with the
target molecule,
as well as enzyme-linked assays which rely on detecting an enzymatic activity
associated with
the target molecule.
Agents that modulate one of the transporters of the present invention can be
identified
using one or more of the above assays, alone or in combination. It is
generally preferable to use
27
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WO 02/083900 PCT/US02/05520
a cell-based or cell free system first and then confirm activity in an animal
or other model
system. Such model systems are well known in the art and can readily be
employed in this
context.
Modulators of transporter protein activity identified according to these drug
screening
assays can be used to treat a subject with a disorder mediated by the
transporter pathway, by
treating cells or tissues that express the transporter. Experimental data as
provided in Figure 1
indicates expression in brain and muscle. These methods of treatment include
the steps of
administering a modulator of transporter activity in a pharmaceutical
composition to a subject
in need of such treatment, the modulator being identified as described herein.
1.0 In yet another aspect of the invention, the transporter proteins can be
used as "bait
proteins" in a two-hybrid assay or three-hybrid assay (see, e.g.~ U.S. Patent
No. 5,283,317;
Zervos et al. (1993) Cell 72:223-232; Madura et al. (1993) J. Biol. Chem.
268:12046-12054;
Bartel et al. (1993) Biotechnigues 14:920-924; Iwabuchi et al. (1993) Oncogene
8:1693-
1696; and Brent W094/10300), to identify other proteins, which bind to or
interact with the
transporter and are involved in transporter activity. Such transporter-binding
proteins are
also likely to be involved in the propagation of signals by the transporter
proteins or
transporter targets as, for example, downstream elements of a transporter-
mediated signaling
pathway. Alternatively, such transporter-binding proteins are likely to be
transporter
inhibitors.
The two-hybrid system is based on the modular nature of most transcription
factors,
which consist of separable DNA-binding and activation domains. Briefly, the
assay utilizes .
two different DNA constructs. In one construct, the gene that codes for a
transporter protein
is fused to a gene encoding the DNA binding domain of a known transcription
factor (e.g.,
GAL-4). In the other construct, a DNA sequence, from a library of DNA
sequences, that
encodes an unidentified protein ("prey" or "sample") is fused to a gene that
codes for the
activation domain of the known transcription factor. If the "bait" and the
"prey" proteins are
able to interact, i~c vivo, forming a transporter-dependent complex, the DNA-
binding and
activation domains of the transcription factor are brought into close
proximity. This
proximity allows transcription of a reporter gene (e.g., LacZ) which is
operably linked to a
transcriptional regulatory site responsive to the transcription factor.
Expression of the
reporter gene can be detected and cell colonies containing the functional
transcription factor
can be isolated and used to obtain the cloned gene which encodes the protein
which interacts
with the transporter protein.
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This invention further pertains to novel agents identified by the above-
described
screening assays. Accordingly, it is within the scope of this invention to
further use an agent
identified as described herein in an appropriate animal model. For example, an
agent
identified as described herein (e.g., a transporter-modulating agent, an
antisense transporter
nucleic acid molecule, a transporter-specific antibody, or a transporter-
binding partner) can
be used in an animal or other model to determine the efficacy, toxicity, or
side effects of
treatment with such an agent. Alternatively, an agent identified as described
herein can be
used in an animal or other model to determine the mechanism of action of such
an agent.
Furthermore, this invention pertains to uses of novel agents identified by the
above-
described screening assays for treatments as described herein.
The transporter proteins of the present invention are also useful to provide a
target for
diagnosing a disease or predisposition to disease mediated by the peptide.
Accordingly, the
invention provides methods for detecting the presence, or levels of, the
protein (or encoding
mRNA) in a cell,.tissue, or organism. Experimental data as provided in Figure
1 indicates
expression in brain and muscle. The method involves contacting a biological
sample with a
compound capable of interacting with the transporter protein such that the
interaction can be
detected. Such an assay can be provided in a single detection format or a
multi-detection
format such. as an antibody chip array.
One agent for detecting a protein in a sample is an antibody capable of
selectively
binding to protein. A biological sample includes tissues, cells and biological
fluids isolated
from a subject, as well as tissues, cells and fluids present within a subject.
The peptides of the present invention also provide targets for diagnosing
active protein
activity, disease, or predisposition to disease, in a patient having a variant
peptide, particularly
activities and conditions that are known for other members of the family of
proteins to which
the present one belongs. Thus, the peptide can be isolated from a biological
sample and
assayed for the presence of a genetic mutation that results in aberrant
peptide. This includes
amino acid substitution, deletion, insertion, rearrangement, (as the result of
aberrant splicing
events), and inappropriate post-translational modification. Analytic methods
include altered
electrophoretic mobility, altered tryptic peptide digest, altered transporter
activity in cell-based
or cell-free assay, alteration in ligand or antibody-binding pattern, altered
isoelectric point,
direct amino acid sequencing, and any other of the known assay techniques
useful for detecting
mutations in a protein. Such an assay can be provided in a single detection
format or a multi-
detection format such as an antibody chip array.
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Ih vitro techniques for detection of peptide include enzyme linked
immunosorbent
assays (ELISAs), Western blots, immunoprecipitations and immunofluorescence
using a
detection reagent, such as an antibody or protein binding agent.
Alternatively, the peptide can
be detected in vivo in a subject by introducing into the subject a labeled
anti-peptide antibody or
other types of detection agent. For example, the antibody can be labeled with
a radioactive
marker whose presence and location in a subject can be detected by standard
imaging
techniques. Particularly useful are methods that detect the allelic variant of
a peptide expressed
in a subject and methods which detect fragments of a peptide in a sample.
The peptides are also useful in pharmacogenomic analysis. Pharmacogenomics
deal
with clinically significant hereditary variations in the response to drugs due
to altered drug
disposition and abnormal action in affected persons. See, e.g., Eichelbaum, M.
(Clip. Exp.
Pha~macol. Physiol. 23(10-11):983-985 (1996)), and Linder, M.W. (Clin. Chem.
43(2):254-266
(1997)). The clinical outcomes of these variations result in severe toxicity
of therapeutic drugs
in certain individuals or therapeutic failure of drugs in certain individuals
as a result of
individual variation in metabolism. Thus, the genotype of the individual can
determine the way
a therapeutic compound acts on the body or the way the body metabolizes the
compound.
Further, the activity of drug metabolizing enzymes effects both the intensity
and duration of
drug action. Thus, the pharmacogenomics of the individual permit the selection
of effective
compounds and effective dosages of such compounds for prophylactic or
therapeutic treatment
based on the individual's genotype. The discovery of genetic polymorphisms in
some drug
metabolizing enzymes has explained why some patients do not obtain the
expected drug effects,
show an exaggerated drug effect, or experience serious toxicity from standard
drug dosages.
Polymorphisms can be expressed in the phenotype of the extensive metabolizer
and the
phenotype of the poor metabolizer. Accordingly, genetic polymorphism may lead
to allelic
protein variants of the transporter protein in which one or more of the
transporter functions in
one population is different from those in another population. The peptides
thus allow a target to
ascertain a genetic predisposition that can affect treatment modality. Thus,
in a ligand-based
. treatment, polymorphism may give rise to amino terminal extracellular
domains and/or other
ligand-binding regions that are more or less active in ligand binding, and
transporter activation.
Accordingly, ligand dosage would necessarily be modified to maximize the
therapeutic effect
within a given population containing a polymorphism. As an alternative to
genotyping, specific
polymorphic peptides could be identified.
CA 02439401 2003-08-25
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The peptides are also useful for treating a disorder characterized by an
absence of,
inappropriate, or unwanted expression of the protein. Experimental data as
provided in Figure
1 indicates expression in brain and muscle. Accordingly, methods for treatment
include the use
of the transporter protein or fragments.
Antibodies
The invention also provides antibodies that selectively bind to one of the
peptides of the
present invention, a protein comprising such a peptide, as well as variants
and fragments
thereof. As used herein, an antibody selectively binds a target peptide when
it binds the target
peptide and does not significantly bind to unrelated proteins. An antibody is
still considered to
selectively bind a peptide even if it also binds to other proteins that are
not substantially
homologous with the target peptide so long as such proteins share homology
with a fragment or
domain of the peptide target of the antibody. In this case, it would be
understood that antibody
binding to the peptide is still selective despite some degree of cross-
reactivity.
As used herein, an antibody is defined in terms consistent with that
recognized within
the art: they are mufti-subunit proteins produced by a mammalian organism in
response to an
antigen challenge. The antibodies of the present invention include polyclonal
antibodies and
monoclonal antibodies, as well as fragments of such antibodies, including, but
not limited to,
Fab or F(ab')2, and Fv fragments.
Many methods are known for generating and/or identifying antibodies to a given
target
peptide. Several such methods are described by Harlow, Antibodies, Cold Spring
Harbor Press,
(1989).
In general, to generate antibodies, an isolated peptide is used as an
immunogen and is
administered to a mammalian organism, such as a rat, rabbit or mouse. The full-
length protein,
an antigenic peptide fragment or a fusion protein can be used. Particularly
important fragments
are those covering functional domains, such as the domains identified in
Figure 2, and domain
of sequence homology or divergence amongst the family, such as those that can
readily be
identified using protein alignment methods and as presented in the Figures.
Antibodies are preferably prepared from regions or discrete fragments of the
transporter proteins. Antibodies can be prepared from any region of the
peptide as described
herein. However, preferred regions will include those involved in
function/activity and/or
transporter/binding partner interaction. Figure 2 can be used to identify
particularly
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CA 02439401 2003-08-25
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important regions while sequence alignment can be used to identify conserved
and unique
sequence fragments.
An antigenic fragment will typically comprise at least 8 contiguous amino acid
residues.
The antigenic peptide can comprise, however, at least 10, 12, 14, 16 or more
amino acid
residues. Such fragments can be selected on a physical property, such as
fragments correspond
to regions that are located on the surface of the protein, e.g., hydrophilic
regions or can be
selected based on sequence uniqueness (see Figure 2).
Detection on an antibody of the present invention can be facilitated by
coupling (i.e.,
physically linking) the antibody to a detectable substance. Examples of
detectable substances
I O include various enzymes, prosthetic groups, fluorescent materials,
luminescent materials,
bioluminescent materials, and radioactive materials. Examples of suitable
enzymes include
horseradish peroxidase, alkaline phosphatase, (3-galactosidase, or
acetylcholinesterase;
examples of suitable prosthetic group complexes include streptavidin/biotin
and avidin/biotin;
examples of suitable fluorescent materials include umbelliferone, fluorescein,
fluorescein
isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride
or
phycoerythrin; an example of a luminescent material includes luminol; examples
of
bioluminescent materials include luciferase, luciferin, and aequorin, and
examples of suitable
radioactive material include l2sI,1311, 3sS or 3H.
Antibody Uses
The antibodies can be used to isolate one of the proteins of the present
invention by
standard techniques, such as affinity chromatography or immunoprecipitation.
The antibodies
can facilitate the purification of the natural protein from cells and
recombinantly produced
protein expressed in host cells. In addition, such antibodies are useful to
detect the presence of
one of the proteins of the present invention in cells or tissues to determine
the pattern of
expression of the protein among various tissues in an organism and over the
course of normal
development. Experimental data as provided in Figure 1 indicates that the
transporter
proteins of the present invention axe expressed in brain and muscle, as
indicated by virtual
northern blot analysis. Further, such antibodies can be used to detect protein
in situ, in vitro, or
in a cell lysate or supernatant in order to evaluate the abundance and pattern
of expression.
Also, such antibodies can be used to assess abnormal tissue distribution or
abnormal expression
during development or progression of a biological condition. Antibody
detection of circulating
fragments of the full length protein can be used to identify turnover.
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Further, the antibodies can be used to assess expression in disease states
such as in
active stages of the disease or in an individual with a predisposition toward
disease related to
the protein's function. When a disorder is caused by an inappropriate tissue
distribution,
developmental expression, level of expression of the protein, or
expressed/processed form, the
antibody can be prepared against the normal protein. Experimental data as
provided in Figure 1
indicates expression in brain and muscle. If a disorder is characterized by a
specific mutation in
the protein, antibodies specific for this mutant protein can be used to assay
for the presence of
the specific mutant protein.
The antibodies can also be used to assess normal and aberrant subcellular
localization of
cells in the various tissues in an organism. Experimental data as provided in
Figure 1 indicates
expression in brain and muscle. The diagnostic uses can be applied, not only
in genetic testing,
but also in monitoring a treatment modality. Accordingly, where treatment is
ultimately aimed
at correcting expression level or the presence of aberrant sequence and
aberrant tissue
distribution or developmental expression, antibodies directed against the
protein or relevant
fragments can be used to monitor therapeutic efFcacy.
Additionally, antibodies are useful in pharmacogenomic analysis. Thus,
antibodies
prepared against polymorphic proteins can be used to identify individuals that
require modified
treatment modalities. °The antibodies are also useful as diagnostic
tools as an immunological
marker for aberrant protein analyzed by electrophoretic mobility, isoelectric
point, tryptic
peptide digest, and other physical assays known to those in the art.
The antibodies are also useful for tissue typing. Experimental data as
provided in
Figure 1 indicates expression in brain and muscle. Thus, where a specific
protein has been
correlated with expression in a specific tissue, antibodies that are specific
for this protein can be
used to identify a tissue type.
The antibodies are also useful for inhibiting protein function, for example,
blocking the
binding of the transporter peptide to a binding partner such as a ligand or
protein binding
partner. These uses can also be applied in a therapeutic context in which
treatment involves
inhibiting the protein's function. An antibody can be used, for example, to
block binding, thus
modulating (agonizing or antagonizing) the peptides activity. Antibodies can
be prepared
against specific fragments containing sites required for function or against
intact protein that is
associated with a cell or cell membrane. See Figure 2 for structural
information relating to the
proteins of the present invention.
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The invention also encompasses kits for using antibodies to detect the
presence of a
protein in a biological sample. The kit can comprise antibodies such as a
labeled or labelable
antibody and a compound or agent for detecting protein in a biological sample;
means for
determining the amount of protein in the sample; means fox comparing the
amount of protein in
the sample with a standard; and instructions for use. Such a kit can be
supplied to detect a single
protein or epitope or can be configured to detect one of a multitude of
epitopes, such as in an
antibody detection array. Arrays are described in detail below for nucleic
acid arrays and
similar methods have been developed for antibody arrays.
' Nucleic Acid Molecules
The present invention further provides isolated nucleic acid molecules that
encode a
transporter peptide or protein of the present invention (cDNA, transcript and
genomic
sequence). Such nucleic acid molecules will consist of, consist essentially
of, or comprise a
nucleotide sequence that encodes one of the transporter peptides of the
present invention, an
allelic variant thereof, or an ortholog or paralog thereof.
As used herein, an "isolated" nucleic acid molecule is one that is separated
from other
nucleic acid present in the natural source of the nucleic acid. Preferably, an
"isolated" nucleic
acid is free of sequences that naturally flank the nucleic acid (i.e.,
sequences located at the 5'
and 3' ends of the nucleic acid) in the genomic DNA of the organism from which
the nucleic
acid is derived. However, there can be some flanking nucleotide sequences, for
example up to
about SKB, 4KB, 3I~B, 2I~B, or II~B or less, particularly contiguous peptide
encoding
sequences and peptide encoding sequences within the same gene but separated by
introns in the
genomic sequence. The important point is that the nucleic acid is isolated
from remote and
unimportant flanking sequences such that it can be subjected to the specific
manipulations
described herein such as recombinant expression, preparation of probes and
primers, and other
uses specific to the nucleic acid sequences.
Moreover, an "isolated" nucleic acid molecule, such as a transcript/cDNA
molecule, can
be substantially free of other cellular material, or culture medium when
produced by
recombinant techniques, or chemical precursors or other chemicals when
chemically
synthesized. However, the nucleic acid molecule can be fused to other coding
or regulatory
sequences and still be considered isolated.
For example, recombinant DNA molecules contained in a vector are considered
isolated. Further examples of isolated DNA molecules include recombinant DNA
molecules
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CA 02439401 2003-08-25
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maintained in heterologous host cells or purified (partially or substantially)
DNA molecules in
solution. Isolated RNA molecules include ih vivo or in vitro RNA transcripts
of the isolated
DNA molecules of the present invention. Isolated nucleic acid molecules
according to the
present invention further include such molecules produced synthetically.
' Accordingly, the present invention provides nucleic acid molecules that
consist of the
nucleotide sequence shown in Figure 1 or 3 (SEQ ID NO:l, transcript sequence
and SEQ ID
N0:3, genomic sequence), or any nucleic acid molecule that encodes the protein
provided in
Figure 2, SEQ ID N0:2. A nucleic acid molecule consists of a nucleotide
sequence when the
nucleotide sequence is the complete nucleotide sequence of the nucleic acid
molecule.
The present invention further provides nucleic acid molecules that consist
essentially of
the nucleotide sequence shown in Figure 1 ox 3 (SEQ ID NO:1, transcript
sequence and SEQ ~
N0:3, genomic sequence), or any nucleic acid molecule that encodes the protein
provided in
Figure 2, SEQ ID N0:2. A nucleic acid molecule consists essentially of a
nucleotide sequence
when such a nucleotide sequence is present with only a few additional nucleic
acid residues in
1 S the final nucleic acid molecule.
The present invention further provides nucleic acid molecules that comprise
the
nucleotide sequences shown in Figure 1 or 3 (SEQ ID NO:1, transcript sequence
and SEQ ID
N0:3, genomic sequence), or any nucleic acid molecule that encodes the protein
provided in
Figure 2, SEQ ID NO:2. A nucleic acid molecule comprises a nucleotide sequence
when the
nucleotide sequence is at least part of the final nucleotide sequence of the
nucleic acid
molecule. In such a fashion, the nucleic acid molecule can be only the
nucleotide sequence or
have additional nucleic acid residues, such as nucleic acid residues that are
naturally associated
with it or heterologous nucleotide sequences. Such a nucleic acid molecule can
have a few
additional nucleotides or can comprise several hundred or more additional
nucleotides. A brief
description of how various types of these nucleic acid molecules can be
readily made/isolated is
provided below.
In Figures 1 and 3, both coding and non-coding sequences are provided. Because
of
the source of the present invention, humans genomic sequence (Figure 3) and
cDNA/transcript sequences (Figure 1), the nucleic acid molecules in the
Figures will contain
genomic intronic sequences, 5' and 3' non-coding sequences, gene regulatory
regions and
non-coding intergenic sequences. In general such sequence features are either
noted in
Figures 1 and 3 or can readily be identified using computational tools known
in the art. As
discussed below, some of the non-coding regions, particularly gene regulatory
elements such
CA 02439401 2003-08-25
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as promoters, are useful for a variety of purposes, e.g. control of
heterologous gene
expression, target for identifying gene activity modulating compounds, and are
particularly
claimed as fragments of the genomic sequence provided herein.
The isolated nucleic acid molecules can encode the mature protein plus
additional
amino or carboxyl-terminal amino acids, or amino acids interior to the mature
peptide (when
the mature form has more than one peptide chain, for instance). Such sequences
may play a
role in processing of a protein from precursor to a mature fonn,,facilitate
protein trafficking,
prolong or shorten protein half life or facilitate manipulation of a protein
for assay or
production, among other things. As generally is the case iu situ, the
additional amino acids may
be processed away from the mature protein by cellular enzymes.
As mentioned above, the isolated nucleic acid molecules include, but are not
limited to,
the sequence encoding the transporter peptide alone, the sequence encoding the
mature peptide
and additional coding sequences, such as a leader or.secretory sequence (e.g.,
a pre-pro or pro-
protein sequence), the sequence encoding the mature peptide, with or without
the additional
coding sequences, plus additional non-coding sequences, for example introns
and non-coding 5'
and 3' sequences such as transcribed but non-translated sequences that play a
role in
transcription, mRNA processing (including splicing and polyadenylation
signals), ribosome
binding and stability of mRNA. In addition, the nucleic acid molecule may be
fused to a
marker sequence encoding, for example, a peptide that facilitates
purification.
Isolated nucleic acid molecules can be in the form of RNA, such as mRNA, or in
the
form DNA, including cDNA and genomic DNA obtained by cloning or produced by
chemical
synthetic techniques or by a combination thereof. The nucleic acid, especially
DNA, can be
double-stranded or single-stranded. Single-stranded nucleic acid can be the
coding strand
(sense strand) or the non-coding strand (anti-sense strand).
~ The invention further provides nucleic acid molecules that encode fragments
of the
peptides of the present invention as well as nucleic acid molecules that
encode obvious variants
of the transporter proteins of the present invention that are described above.
Such nucleic acid
molecules may be naturally occurring, such as allelic variants (same locus),
paralogs (different
locus), and orthologs (different organism), or may be constructed by
recombinant DNA
methods or by chemical synthesis. Such non-naturally occurring variants may be
made by
mutagenesis techniques, including those applied to nucleic acid molecules,
cells, or organisms.
Accordingly, as discussed above, the variants can contain nucleotide
substitutions, deletions,
inversions and insertions. Variation can occur in either or both the coding
and non-coding
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regions. The variations can produce both conservative and non-conservative
amino acid
substitutions.
The present invention further provides non-coding fragments of the nucleic
acid
molecules provided in Figures 1 and 3. Preferred non-coding fragments include,
but are not
limited to, promoter sequences, enhancer sequences, gene modulating sequences
and gene
termination sequences. Such fragments are useful in controlling heterologous
gene expression
and in developing screens to identify gene-modulating agents. A promoter can
readily be
identified as being 5' to the ATG start site in the genomic sequence provided
in Figure 3.
A fragment comprises a contiguous nucleotide sequence greater than 12 or more
nucleotides. Further, a fragment could at least 30, 40, 50,100, 250 or 500
nucleotides in length.
The length of the fragment will be based on its intended use. For example, the
fragment can
encode epitope bearing regions of the peptide, or can be useful as DNA probes
and primers.
Such fragments can be isolated using the known nucleotide sequence to
synthesize an
oligonucleotide probe. A labeled probe can then be used to screen a cDNA
library, genomic
DNA library, or mRNA to isolate nucleic acid corresponding to the coding
region. Further,
primers can be used in PCR reactions to clone specific regions of gene.
A probe/primer typically comprises substantially a purified oligonucleotide or
oligonucleotide pair. The oligonucleotide typically comprises a region of
nucleotide sequence
that hybridizes under stringent conditions to at least about 12, 20, 25, 40,
50 or more
consecutive nucleotides.
Orthologs, homologs, and allelic variants can be identified using methods well
known
in the art. As described in the Peptide Section, these variants comprise a
nucleotide sequence
encoding a peptide that is typically 60-70%, 70-80%, 80-90%, and more
typically at least about
90-95% or more homologous to the nucleotide sequence shown in the Figure
sheets or a
fragment of this sequence. Such nucleic acid molecules can readily be
identified as being able
to hybridize under moderate to stringent conditions, to the nucleotide
sequence shown in the
Figure sheets or a fragment of the sequence. Allelic variants can readily be
determined by
genetic locus of the encoding gene. The gene encoding the novel transporter
protein of the
present invention is located on a genome component that has been mapped to
human
chromosome 5 (as indicated in Figure 3), which is supported by multiple lines
of evidence, such
as STS and BAC map data.
Figure 3 provides information on SNPs that have been found in the gene
encoding
the transporter protein of the present invention. SNPs-were identified at 269
different
37
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nucleotide positions, including non-synonymous coding SNPs at positions
166328, 169076,
171899, and 171919. Changes in the amino acid sequence caused by these SNPs is
indicated
in Figure 3 and can readily be determined using the universal genetic code and
the protein
sequence provided in Figure 2 as a reference. Some of the SNPs that are
located outside the
ORF and in introns may affect gene transcription.
As used herein, the term "hybridizes under stringent conditions" is intended
to describe
conditions for hybridization and washing under which nucleotide sequences
encoding a peptide
at least 60-70% homologous to each other typically remain hybridized to each
other. The
conditions can be such that sequences at least about 60%, at least about 70%,
or at least about
80% or more homologous to each other typically remain hybridized to each
other. Such
stringent conditions are known to those skilled in the art and can be found in
Current Protocols
in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. One example
of stringent
hybridization conditions are hybridization in 6X sodium chloride/sodium
citrate (SSC) at about
45C, followed by one or more washes in 0.2 X SSC, 0.1% SDS at 50-65C. Examples
of
moderate to low stringency hybridization conditions are well known in the art.
Nucleic Acid Molecule Uses
The nucleic acid molecules of the present invention are useful for probes,
primers,
chemical intermediates, and in biological assays. The nucleic acid molecules
are useful as a
hybridization probe for messenger RNA, transcript/cDNA and genomic DNA to
isolate full-
length cDNA and genomic clones encoding the peptide described in Figure 2 and
to isolate
cDNA and genomic clones that correspond to variants (alleles, orthologs, etc.)
producing the
same or related peptides shown in Figure 2. As illustrated in Figure 3, SNPs
were identified at
269 different nucleotide positions.
The probe can correspond to any sequence along the entire length of the
nucleic acid
molecules provided in the Figures. Accordingly, it could be derived from 5'
noncoding
regions, the coding region, and 3' noncoding regions. However, as discussed,
fragments are not
to be construed as encompassing fragments disclosed prior to the present
invention.
The nucleic acid molecules are also useful as primers for PCR to amplify any
given
region of a nucleic acid molecule and are useful to synthesize antisense
molecules of desired
length and sequence.
The nucleic acid molecules are also useful for constructing recombinant
vectors. Such
vectors include expression vectors that express a portion of, or all of, the
peptide sequences.
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Vectors also include insertion vectors, used to integrate into another nucleic
acid molecule
sequence, such as into the cellular genome, to alter ih situ expression of a
gene and/or gene
product. For example, an endogenous coding sequence can be replaced via
homologous
recombination with all or part of the coding region containing one or more
specifically
introduced mutations.
The nucleic acid molecules are also useful for expressing antigenic portions
of the
proteins.
The nucleic acid molecules are also useful as probes for determining the
chromosomal
positions of the nucleic acid molecules by means of in situ hybridization
methods. The gene
I O encoding the novel transporter protein of the present invention is located
on a genome
component that has been mapped to human chromosome 5 (as indicated in Figure
3), which is
supported by multiple lines of evidence, such as STS and BAC map data.
The nucleic acid molecules are also useful in making vectors containing the
gene
regulatory regions of the nucleic acid molecules of the present invention.
The nucleic acid molecules are also useful for designing ribozymes
corresponding to all,
or a part, of the mRNA produced from the nucleic acid molecules described
herein.
The nucleic acid molecules are also useful for making vectors that express
part, or all, of
the peptides.
'The nucleic acid molecules are also useful for constructing host cells
expressing a part,
or all, of the nucleic acid molecules and peptides.
The nucleic acid molecules are also useful for constructing transgenic animals
expressing all, or a part, of the nucleic acid molecules and peptides.
The nucleic acid molecules are also useful as hybridization probes for
determining the
presence, level, form and distribution of nucleic acid expression.
Experimental data as
provided in Figure 1 indicates that the transporter proteins of the present
invention are
expressed in brain and muscle, as indicated by virtual northern blot analysis.
Accordingly, the probes can be used to detect the presence of, or to determine
levels of,
a specific nucleic acid molecule in cells, tissues, and in organisms. The
nucleic acid whose
level is determined can be DNA or RNA. Accordingly, probes corresponding to
the peptides
3 0 described herein can be used to assess expression and/or gene copy-number
in a given cell,
tissue, or organism. These uses are relevant for diagnosis of disorders
involving an increase or
decrease in transporter protein expression relative to normal results.
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In vitro techniques for detection of mRNA include Northern hybridizations and
ih situ
hybridizations. In vitro techniques for detecting DNA include Southern
hybridizations and in
situ hybridization.
Probes can be used as a part of a diagnostic test kit for identifying cells or
tissues that
express a transporter protein, such as by measuring a level of a transporter-
encoding nucleic
acid in a sample of cells from a subject e.g., mRNA or genomic DNA, or
determining if a
transporter gene has been mutated. Experimental data as provided in Figure 1
indicates that
the transporter proteins of the present invention are expressed in brain and
muscle, as
indicated by virtual northern blot analysis.
Nucleic acid expression assays are useful for drug screening to identify
compounds that
modulate transporter nucleic acid expression.
'The invention thus provides a method for identifying a compound that can be
used to
treat a disorder associated with nucleic acid expression of the transporter
gene, particularly
biological and pathological processes that are mediated by the transporter in
cells and tissues
that express it. Experimental data as provided in Figure 1 indicates
expression in brain and
muscle. The method typically includes assaying the ability of the compound to
modulate the
expression of the transporter nucleic acid and thus identifying a compound
that can be used to
treat a disorder characterized by undesired transporter nucleic acid
expression. The assays can
be performed in cell-based and cell-free systems. Cell-based assays include
cells naturally
expressing the transporter nucleic acid or recombinant cells genetically
engineered to express
specific nucleic acid sequences.
The assay for transporter nucleic acid expression can involve direct assay of
nucleic
acid levels, such as mRNA levels, or on collateral compounds involved in the
signal pathway.
Further, the expression of genes that are up- or down-regulated in response to
the transporter
protein signal pathway can also be assayed. In this embodiment the regulatory
regions of these
genes can be operably linked to a reporter gene such as luciferase.
Thus, modulators of transporter gene expression can be identified in a method
wherein a
cell is contacted with a candidate compound and the expression of mRNA
determined. The
level of expression of transporter mRNA in the presence of the candidate
compound is
compared to the level of expression of transporter mRNA in the absence of the
candidate
compound. The candidate compound can then be identified as a modulator of
nucleic acid
expression based on this comparison and be used, for example to treat a
disorder characterized
by aberrant nucleic acid expression. When expression of mRNA is statistically
significantly
CA 02439401 2003-08-25
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greater in the presence of the candidate compound than in its absence, the
candidate compound
is identified as a stimulator of nucleic acid expression. When nucleic acid
expression is
statistically significantly less in the presence of the candidate compound
than in its absence, the
candidate compound is identified as an inhibitor of nucleic acid expression.
The invention further provides methods of treatment, with the nucleic acid as
a target,
using a compound identified through drug screening as a gene modulator to
modulate
transporter nucleic acid expression in cells and tissues that express the
transporter.
Experimental data as provided in Figure 1 indicates that the transporter
proteins of the
present invention are expressed in brain and muscle, as indicated by virtual
northern blot
analysis. Modulation includes both up-regulation (i.e. activation or
agonization) or down-
regulation (suppression or antagonization) or nucleic acid expression.
Alternatively, a modulator for txansporter nucleic acid expression can be a
small
molecule or drug identified using the screening assays described herein as
long as the drug or
small molecule inhibits the transporter nucleic acid expression in the cells
and tissues that
express the protein. Experimental data as provided in Figure 1 indicates
expression in brain and
muscle.
The nucleic acid molecules are also useful for monitoring the effectiveness of
modulating compounds on the expression or activity of the transporter gene in
clinical trials or
in a treatment regimen. Thus, the gene expression pattern can serve as a
barometer for the
continuing effectiveness of treatment with the compound, particularly with
compounds to
which a patient can develop resistance. The gene expression pattern can also
serve as a marker
indicative of a physiological response of the affected cells to the compound.
Accordingly, such
monitoring would allow either increased administration of the compound or the
administration
of alternative compounds to which the patient has not become resistant.
Similarly, if the level
of nucleic acid expression falls below a desirable level, administration of
the compound could
be commensurately decreased.
The nucleic acid molecules are also useful in diagnostic assays for
qualitative changes
in transporter nucleic acid expression, and particularly in qualitative
changes that lead to
pathology. The nucleic acid molecules can be used to detect mutations in
transporter genes and
gene expression products such as mRNA. The nucleic acid molecules can be used
as
hybridization probes to detect naturally occurring genetic mutations in the
transporter gene and
thereby to determine whether a subject with the mutation is at risk for a
disorder caused by the
mutation. Mutations include deletion, addition, or substitution of one or more
nucleotides in the
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gene, chromosomal rearrangement, such as inversion or transposition,
modification of genomic
DNA, such as aberrant methylation patterns or changes in gene copy number,
such as
amplification. Detection of a mutated form of the transporter gene associated
with a
dysfunction provides a diagnostic tool for an active disease or susceptibility
to disease when the
disease results from overexpression, underexpression, or altered expression of
a transporter
protein.
Individuals carrying mutations in the transporter gene can be detected at the
nucleic acid
Level by a variety of techniques. Figure 3 provides information on SNPs that
have been found
in the gene encoding the transporter protein of the present invention. SNPs
were identified at
269 different nucleotide positions, including non-synonymous coding SNPs at
positions
166328, 169076, 171899, and 171919. Changes in the amino acid sequence caused
by these
SNPs is indicated in Figure 3 and can readily be determined using the
universal genetic code
and the protein sequence provided in Figure 2 as a reference. Some of the SNPs
that are
located outside the ORF and in introns may affect gene transcription. The gene
encoding the
novel transporter protein of the present invention is located on a genome
component that has
been mapped to human chromosome 5 (as indicated in Figure 3), which is
supported by
multiple lines of evidence, such as STS and BAC map data. Genomic DNA can be
analyzed
directly or can be amplified by using PCR prior to analysis. RNA or cDNA can
be used in the
same way. In some uses, detection of the mutation involves the use of a
probe/primer in a
polymerase chain reaction (PCR) (see, e.g. U.S. Patent Nos. 4,683,195 and
4,683,202), such as
anchor PCR or RACE PCR, or, alternatively, in a ligation chain reaction (LCR)
(see, e.g.,
Landegran et al., Science 241:1077-1080 (1988); and Nakazawa et al., PNAS
91:360-364
(1994)), the latter of which can be particularly useful for detecting point
mutations in the gene
(see Abravaya et al., Nucleic Acids Res. 23:675-682 (1995)). This method can
include the steps
of collecting a sample of cells from a patient, isolating nucleic acid (e.g.,
genomic, mRNA or
both) from the cells of the sample, contacting the nucleic acid sample with
one or more primers
which specifically hybridize to a gene under conditions such that
hybridization and
amplification of the gene (if present) occurs, and detecting the presence or
absence of an
amplification product, or detecting the size of the amplification product and
comparing the
length to a control sample. Deletions and insertions can be detected by a
change in size of the
amplified product compared to the normal genotype. Point mutations can be
identified by
hybridizing amplified DNA to normal RNA or antisense DNA sequences.
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Alternatively, mutations in a transporter gene can be directly identified, for
example, by
alterations in restriction enzyme digestion patterns determined by gel
electrophoresis.
Further, sequence-specific ribozymes (U.S. Patent No. 5,498,531) can be used
to score
for the presence of specific mutations by development or loss of a ribozyme
cleavage site.
Perfectly matched sequences can be distinguished from mismatched sequences by
nuclease
cleavage digestion assays or by differences in melting temperature.
Sequence changes at specific locations can also be assessed by nuclease
protection
assays such as RNase and S 1 protection or the chemical cleavage method.
Furthermore,
sequence differences between a mutant transporter gene and a wild-type gene
can be
determined by direct DNA sequencing. A variety of automated sequencing
procedures can be
wtilized when performing the diagnostic assays (Naeve, C.W., (1995)
Biotechv~iques 19:448);
including sequencing by mass spectrometry (see; e.g., PCT International
Publication No. WO
94/16101; Cohen et al., Adv. Chromatogr. 36:127-162 (1996); and Griffin et
al., Appl.
Biochem. Biotechnol. 38:147-159 (1993)}. . .
Other methods for detecting mutations in the gene include methods in which
protection
from cleavage agents is used to detect mismatched bases in RNA/RNA or RNA/DNA
duplexes
(Myers et al., Science 230:1242 (1985)); Cotton et al., PNAS 85:4397 (1988);
Saleeba et al.,
Meth. Enzymol. 217:286-295 (1992)), electrophoretic mobility of mutant and
wild type nucleic
acid is compared (Orita et al., PNAS 86:2766 (1989); Cotton et al., Mutat.
Res. 285:125-144
(1993); and Hayashi et al., Genet. Anal. Tech. Appl. .9:73-79 (1992)), and
movement of mutant
or wild-type fragments in polyacrylamide gels containing a gradient of
denaturant is assayed
using denaturing gradient gel electrophoresis (Myers et al., Nature 313:495
(1985)). Examples
of other techniques for detecting point mutations include selective
oligonucleotide
hybridization, selective amplification, and selective primer extension.
The nucleic acid molecules are also useful for testing an individual for a
genotype that
while not necessarily causing the disease, nevertheless affects the treatment
modality. Thus, the
nucleic acid molecules can be used to study the relationship between an
individual's genotype
and the individual's response to a compound used for treatment
(pharmacogenomic
relationship). Accordingly, the nucleic acid molecules described herein can be
used to assess
the mutation content of the transporter gene in an individual in order to
select an appropriate
compound or dosage regimen for treatment. Figure 3 provides information on
SNPs that have
been found in the gene encoding the transporter protein of the present
invention. SNPs were
identified at 269 different nucleotide positions, including non-synonymous
coding SNPs at
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positions 166328, 169076, 171899, and 171919. Changes in the amino acid
sequence caused
by these SNPs is indicated in Figure 3 and can readily be determined using the
universal
genetic code and the protein sequence provided in Figure 2 as a reference.
Some of the SNPs
that are located outside the ORF and in introns may affect gene transcription.
Thus nucleic acid molecules displaying genetic variations that affect
treatment provide a
diagnostic target that can be used to tailor treatment in an individual.
Accordingly, the
production of recombinant cells and animals containing these polymorphisms
allow effective
clinical design of treatment compounds and dosage regimens.
The nucleic acid molecules are thus useful as antisense constructs to confirol
transporter
gene expression in cells, tissues, and organisms. A DNA antisense nucleic acid
molecule is
designed to be complementary to a region of the gene involved in
transcription, preventing
transcription and hence production of transporter protein. An antisense RNA or
DNA nucleic
acid molecule would hybridize to the mRNA and thus block translation of mRNA
into
transporter protein.
Alternatively, a class of antisense molecules can be used to inactivate mRNA
in order to
decrease expression of transporter nucleic acid. Accordingly, these molecules
can treat a
disorder characterized by abnormal or undesired transporter nucleic acid
expression. This
technique involves cleavage by means of ribozymes'containing nucleotide
sequences
complementary to one or more regions in the mRNA that attenuate the ability of
the mRNA to
be translated. Possible regions include coding regions and particularly coding
regions
. corresponding to the catalytic and other functional activities of the
transporter protein, such as
ligand binding. ..
The nucleic acid molecules also provide vectors for gene therapy in patients
containing
cells that are aberrant in transporter gene expression. Thus, recombinant
cells, which include
the patient's cells that have been engineered ex vivo and returned to the
patient, are introduced
into an individual where the cells produce the desired transporter protein to
treat the individual.
The invention also 'encompasses kits for detecting the presence of a
transporter nucleic
acid in a biological sample. Experimental data as provided in Figure 1
indicates that the
transporter proteins of the present invention are expressed in brain and
muscle, as indicated
by virtual northern blot analysis. For example, the kit can comprise reagents
such as a labeled
or labelable nucleic acid or agent capable of detecting transporter nucleic
acid in a biological
sample; means for determining the amount of transporter nucleic acid in the
sample; and means
for comparing the amount of transporter nucleic acid in the sample with a
standard. The
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compound or agent can be packaged in a suitable container. The kit can further
comprise
instructions for using the kit to detect transporter protein mRNA or DNA.
Nucleic Acid Arrays
The present invention further provides nucleic acid detection kits, such as
arrays or
microarrays of nucleic acid molecules that are based on the sequence
information provided
in Figures l and 3 (SEQ ID NOS:1 and 3).
As used herein "Arrays" or "Microarrays" refers to an array of distinct
polynucleotides or oligonucleotides synthesized on a substrate, such as paper,
nylon or other
type of membrane, filter, chip, glass slide, or any other suitable solid
support. In one
embodiment, the microarray is prepared and used according to the methods
described in US
Patent 5,837,832, Chee et al., PCT application W095111995 (Chee et al.),
Lockhart, D. J. et
al. (1996; Nat. Biotech. 14: 1675-1680) and Schena, M. et al. (1996; Proc.
Natl. Acad. Sci.
93: 10614-10619), all of which axe incorporated herein in their entirety by
reference. In
other embodiments, such arrays are produced by the methods described by Brown
et al., US
Patent No. 5,807,522.
The microarray or detection kit is preferably composed of a large number of
unique,
single-stranded nucleic acid sequences, usually either synthetic antisense
oligonucleotides or
fragments of cDNAs, fixed to a solid support. The oligonucleotides axe
preferably about 6-
60 nucleotides in length, more preferably 15-30 nucleotides in length, and
most preferably
about 20-25 nucleotides in length. For a certain type of microarray or
detection kit, it may be
preferable to use oligonucleotides that are only 7-20 nucleotides in length.
The microarray or
detection kit may contain oligonucleotides that.cover the known 5', or 3',
sequence,
sequential oligonucleotides that cover the full length sequence; or unique
oligonucleotides
selected from particular areas along the length of the sequence.
Polynucleotides used in the
microaxray or detection kit may be oligonucleotides that axe specific to a
gene or genes of
interest.
In order to produce oligonucleotides to a known sequence for a microarray or
detection kit, the genes) of interest (or an ORF identified from the contigs
of the present
invention) is typically examined using a computer algorithm which starts at
the 5' or at the 3'
end of the nucleotide sequence. Typical algorithms will then identify
oligomers of defined
length that are unique to the gene, have a GC content within a range suitable
for
hybridization, and lack predicted secondary structure that may interfere with
hybridization.
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In certain situations it may be appropriate to use pairs of oligonucleotides
on a microarray or
detection kit. The "pairs" will be identical, except for one nucleotide that
preferably is
located in the center of the sequence. The second oligonucleotide in the pair
(mismatched by
one) serves as a control. The number of oligonucleotide pairs may range from
two to one
million. The oligomers are synthesized at designated areas on a substrate
using a light-
directed chemical process. The substrate may be paper, nylon or other type of
membrane,
filter, chip, glass slide or any other suitable solid support.
In another aspect, an oligonucleotide may be synthesized on the surface of the
substrate by using a chemical coupling procedure and an ink jet application
apparatus, as
described in PCT application W095/251 I 16 (Baldeschweiler et al. ) which is
incorporated
herein in its entirety by reference. In another aspect, a "gridded" array
analogous to a dot (or
slot) blot may be used to arrange and link cDNA fragments or oligonucleotides
to the surface
of a substrate using a vacuum system, thermal, UV, mechanical or chemical
bonding
procedures. An array, such as those described above, may be produced by hand
or by using
available devices (slot blot or dot blot apparatus), materials (any suitable
solid support), and
machines (including robotic instruments), and may contain 8, 24, 96, 384,
1536, 6144 or
more oligonucleotides, or any other number between two and one million which
lends itself
to the efficient use of commercially available instrumentation.
In order to conduct sample analysis using a microarray or detection kit, the
RNA or
DNA from a biological sample is made into hybridization probes. The mRNA is
isolated,
and cDNA is produced and used as a template to make antisense RNA (a.RNA). The
aRNA
is amplified in the presence of fluorescent nucleotides, and labeled probes
axe incubated with
the microarray or detection kit so that the probe sequences hybridize to
complementary
oligonucleotides of the microarray or detection kit. Incubation conditions are
adjusted so that
hybridization occurs with precise complementary matches or with various
degrees of less
complementarity. After removal of nonhybridized probes, a scanner is used to
determine the
levels and patterns of fluorescence. The scanned images are examined to
determine degree of
complementarity and the relative abundance of each oligonucleotide sequence on
the
microarray or detection kit. The biological samples may be obtained from any
bodily fluids
(such as blood, urine, saliva, phlegm, gastric juices, etc.), cultured cells,
biopsies, or other
tissue preparations. A detection system may be used to measure the absence,
presence, and
amount of hybridization for all of the distinct sequences simultaneously. This
data may be
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used for large-scale correlation studies on the sequences, expression
patterns, mutations,
variants, or polymorphisms among samples.
Using such arrays, the present invention provides methods to identify the
expression
of the transporter proteins/peptides of the present invention. In detail, such
methods
. comprise incubating a test sample with one or more nucleic acid molecules
and assaying for
binding of the nucleic acid molecule with components within the test sample.
Such assays
will typically involve arrays comprising many genes, at least one of which is
a gene of the
present invention and or alleles of the transporter gene of the present
invention. Figure 3
provides information on SNPs that have been found in the gene encoding the
transporter
protein of the present invention. SNPs were identified at 269 different
nucleotide positions,
including non-synonymous coding SNPs at positions 166328, 169076, 171899, and
171919.
Changes in the amino acid sequence caused by these SNPs is indicated in Figure
3 and can
readily be determined using the universal genetic code and the protein
sequence provided in
Figure 2 as,a reference. Some of the SNPs that are located outside the ORF and
in introns
may affect gene transcription.
Conditions for incubating a nucleic acid molecule with a test sample vary.
Incubation conditions depend on the format employed in the assay, the
detection methods
employed, and the type and nature of the nucleic acid molecule used in the
assay. One
skilled in the art will recognize that any one of the commonly available
hybridization,
amplification or array assay formats can readily be adapted to employ the
novel fragments of
the Human genome disclosed herein. Examples of such assays can be found in
Chard, T, An
Introduction to Radioimmunoassay and Related Techniques, Elsevier Science
Publishers,
Amsterdam, The Netherlands (1986); Bullock, G. R. et al., Techniques i~z
Immuhocytochemistry, Academic Press, Orlando, FL Vol. 1 (1 982), Vol. 2
(1983), Vol. 3
(1985); Tijssen, P., Practice and Theory of Enzyme Immunoassays: Laboratory
Techniques
in Biochemistry and Molecular Biology, Elsevier Science Publishers, Amsterdam,
The
Netherlands (1985).
The test samples of the present invention include cells, protein or membrane
extracts
of cells. The test sample used in the above-described method will vary based
on the assay
format, nature of the detection method and the tissues, cells or extracts used
as the sample to
be assayed. Methods for preparing nucleic acid extracts or of cells are well
known in the art
and can be readily be adapted in order to obtain a sample that is compatible
with the system
utilized.
47
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
In another embodiment of the present invention, kits are provided which
contain the
necessary reagents to carry out the assays of the present invention.
Specifically, the invention provides a compartmentalized kit to receive, in
close
confinement, one or more containers which comprises: (a) a first container
comprising one
of the nucleic acid molecules that can bind to a fragment of the Human genome
disclosed
herein; and (b) one or more other containers comprising one or more of the
following: wash
reagents, reagents capable of detecting presence of a bound nucleic acid.
In detail, a compartmentalized kit includes any kit in which reagents are
contained in
separate containers. Such containers include small glass containers, plastic
containers, strips
of plastic, glass or paper, or arraying material such as silica. Such
containers allows one to
efficiently transfer reagents from one compartment to another compartment such
that the
samples and reagents are not cross-contaminated, and the agents or solutions
of each
container can be added in a quantitative fashion from one compartment to
another. Such
containers will include a container which will accept the test sample, a
container which
contains the nucleic acid probe, containers which contain wash reagents (such
as phosphate
buffered saline, Tris-buffers, etc.), and containers which contain the
reagents used to detect
the bound probe. One skilled in the art will readily recognize that the
previously
unidentified transporter gene of the present invention can be routinely
identified using the
sequence information disclosed herein can be readily incorporated into one of
the established
kit formats which are well known in the art, particularly expression arrays.
Vectors/host cells
The invention also provides vectors containing the nucleic acid molecules
described
herein. The term "vector" refers to a vehicle, preferably a nucleic acid
molecule, which can
transport the nucleic acid molecules. When the vector is a nucleic acid
molecule, the nucleic
acid molecules are covalently linked to the vector nucleic acid. With this
aspect of the
invention, the vector includes a plasmid, single or double stranded phage, a
single or double
stranded RNA or DNA viral vector, or artificial chromosome, such as a BAC,
PAC, YAC, OR
MAC.
A vector can be maintained in the host cell as an extrachromosomal element
where it
replicates and produces additional copies of the nucleic acid molecules.
Alternatively, the
vector may integrate into the host cell genorne and produce additional copies
of the nucleic acid
molecules when the host cell replicates.
4~
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The invention provides vectors for the maintenance (cloning vectors) or
vectors for
expression (expression vectors) of the nucleic acid molecules. The vectors can
function in
procaryotic or eukaryotic cells or in both (shuttle vectors).
Expression vectors contain cis-acting regulatory regions that are operably
linked in the
vector to the nucleic acid molecules such that transcription of the nucleic
acid molecules is
allowed in a host cell. The nucleic acid molecules can be introduced into the
host cell with a
separate nucleic acid molecule capable of affecting transcription. Thus, the
second nucleic acid
molecule may provide a trans-acting factor interacting with the cis-regulatory
control region to
allow transcription of the nucleic acid molecules from the vector.
Alternatively, a trans-acting
factor may be supplied by the host cell. Finally, a trans-acting factor can be
produced from the
vector itself. It is understood, however, that in some embodiments,
transcription and/or
translation of the nucleic acid molecules can occur in a cell-free system.
The regulatory sequence to which the nucleic acid molecules described herein
can be
operably linked include promoters for directing mRNA transcription. These
include, but are
not limited to, the left promoter from bacteriophage ~,, the lac, TRP, and TAC
promoters from
E. coli, the early and late promoters from SV40, the CMV immediate early
promoter, the
adenovirus early and late promoters, and retrovirus long-terminal repeats.
In addition to control regions that promote transcription, expression vectors
may also
include regions that modulate transcription, such as repressor binding sites
and enhancers.
Examples include the SV40 enhancer, the cytomegalovirus immediate early
enhancer, polyoma
enhancer, adenovirus enhancers, and retrovirus LTR enhancers.
In addition to containing sites for transcription initiation and control,
expression vectors
can also contain sequences necessary for transcription termination and, in the
transcribed region
a ribosome binding site for translation. Other regulatory control elements for
expression
include initiation and termination codons as well as polyadenylation signals.
The person of
ordinary skill in the art would be aware of the numerous regulatory sequences
that are useful in
expression vectors. Such regulatory sequences are described, for example, in
Sambrook et al.,
Molecular Cloning: A Laboratory Manual. 2nd. ed., Cold Spring Harbor
Laboratory Press,
Cold Spring Harbor, NY, (1989).
A variety of expression vectors can be used to express a nucleic acid
molecule. Such
vectors include chromosomal, episomal, and virus-derived vectors, for example
vectors derived
from bacterial plasmids, from bacteriophage, from yeast episomes, from yeast
chromosomal
elements, including yeast artificial chromosomes, from viruses such as
baculoviruses,
49
CA 02439401 2003-08-25
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papovaviruses such as SV40, Vaccinia viruses, adenoviruses, poxviruses,
pseudorabies viruses,
and retroviruses. Vectors may also be derived from combinations of these
sources such as
those derived from plasmid and bacteriophage genetic elements, e.g. cosmids
and phagemids.
Appropriate cloning and expression vectors for prokaryotic and eukaryotic
hosts are described
in Sambrook et al., Molecular Cloning: A Labo~atoty Manual. 2nd. ed., Cold
Spring Harbor
Laboratory Press, Cold Spring Harbor, NY, (1989).
The regulatory sequence may provide constitutive expression in one or more
host cells
(i-.e. tissue specific) or may provide for inducible expression in one or more
cell types such as
by temperature, nutrient additive, or exogenous factor such as a hormone or
other ligand. A
- variety of vectors providing for constitutive and inducible expression in
prokaryotic and
eukaryotic hosts are well known to those of ordinary skill in the art.
The nucleic acid molecules can be inserted into the vector nucleic acid by
well-known
methodology. Generally, the DNA sequence that will ultimately be expressed is
joined to an
expression vector by cleaving the DNA sequence and the expression vector with
one or more
restriction enzymes and then ligating the fragments together. Procedures for
restriction enzyme
digestion and ligation are well known to those of ordinary skill in the art.
The vector containing the appropriate nucleic acid molecule can be introduced
into an
appropriate host cell for propagation or expression using well-known
techniques. Bacterial
cells include, but are not limited to, E. coli, Streptomyces, and Salmonella
Zyphimurium.
Eukaryotic cells include, but are not limited to, yeast, insect cells such as
Drosophila, animal
cells such as COS and CHO cells, and plant cells.
As described herein, it may be desirable to express the peptide as a fusion
protein.
Accordingly, the invention provides fusion vectors that allow for the
production of the peptides.
Fusion vectors can increase the expression of a recombinant protein, increase
the solubility of
the recombinant protein, and aid in the purification of the protein by acting
for example as a
ligand for affinity purification. A proteolytic cleavage site may be
introduced at the junction of
the fusion moiety so that the desired peptide can ultimately be separated from
the fusion
moiety. Proteolytic enzymes include, but are not limited to, factor Xa,
thrombin, and
enterotransporter. Typical fusion expression vectors include pGEX (Smith et
al., Gene 67:31-
40 (1988)), pMAL (New England Biolabs, Beverly, IvIA) and pRITS (Pharmacia,
Piscataway,
NJ) which fuse glutathione S-transferase (GST), maltose E binding protein, or
protein A,
respectively, to the target recombinant protein. Examples of suitable
inducible non-fusion E.
CA 02439401 2003-08-25
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coli expression vectors include pTrc (Amann et al., Gene 69:301-315 (1988))
and pET 1 1d
(Studier et al., Gene Expression Technology: Methods in Enzymology 185:60-89
(1990)).
Recombinant protein expression can be maximized in host bacteria by providing
a
genetic background wherein the host cell has an impaired capacity to
proteolytically cleave the
recombinant protein. (Gottesman, S., Gene Expression Technology: Methods in
Enzymology
185, Academic Press, San Diego, California (1990) 119-128). Alternatively, the
sequence of
the nucleic acid molecule of interest can be altered to provide preferential
codon usage for a
specific host cell, for example E. coli. (Wada et al., Nucleic Acids Res.
20:2111-2118 (1992)).
The nucleic acid molecules can also be expressed by expression vectors that
are
operative in yeast. Examples of vectors for expression in yeast e.g., S
cere~isiae include
pYepSecl (Baldari, et al., EMBO J. 6:229-234 (1987)), pMFa (Kurjan et al.,
Cell 30:933-
943(1982)), pJRY88 (Schultz et al., Gene 54:113-123 (1987)), and pYES2
(Invitrogen
Corporation, San Diego, CA).
The nucleic acid molecules can also be expressed in insect cells using, for
example,
baculovirus expression vectors. Baculovirus vectors available for expression
of proteins in
cultured insect cells (e.g., Sf 9 cells) include the pAc series (Smith et al.,
Mol. Cell Biol.
3:2156-2165 (1983)) and the pVL series (Lucklow et al., Virology 170:31-39
(1989)).
In certain embodiments of the invention, the nucleic acid molecules described
herein are
expressed in mammalian cells using mammalian expression vectors. Examples of
mammalian
expression vectors include pCDM8 (Seed, B. Nature 329:840(1987)) and pMT2PC
(I~aufinan
et al., EMBO J. 6:187-195 (1987)).
The expression vectors listed herein are provided by way of example only of
the well-
known vectors available to those of ordinary skill in the art that would be
useful to express the
nucleic acid molecules. The person of ordinary skill in the art would be aware
of other vectors
suitable for maintenance propagation or expression of the nucleic acid
molecules described
herein. These are found for example in Sambrook, J., Fritsh, E. F., and
Maniatis, T. Molecular
Cloning: A Laboratory Manual. 2nd, ed., Cold Spring Harbor Laboratory, Cold
Spring Harbor
Laboratory Press, Cold Spring Harbor, NY,1989.
The invention also encompasses vectors in which the nucleic acid sequences
described
herein are cloned into the vector in reverse orientation, but operably linked
to a regulatory
sequence that permits transcription of antisense RNA. Thus, an antisense
transcript can be
produced to all, or to a portion, of the nucleic acid molecule sequences
described herein,
including both coding and non-coding regions. Expression of this antisense RNA
is subject to
51
CA 02439401 2003-08-25
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each of the parameters described above in relation to expression of the sense
RNA (regulatory
sequences, constitutive or inducible expression, tissue-specific expression).
The invention also relates to recombinant host cells containing the vectors
described
herein. Host cells therefore include prokaryotic cells, lower eukaryotic cells
such as yeast, other
eukaryotic cells such as insect cells, and higher eukaryotic cells such as
mammalian cells.
The recombinant host cells are prepared by introducing the vector constructs
described
herein into the cells by techniques readily available to the person of
ordinary skill in the art.
These include, but are not limited to, calcium phosphate transfection, DEAE-
dextran-mediated
transfection, cationic lipid-mediated transfection, electroporation,
transduction, infection,
lipofection, and other techniques such as those found in Sambrook, et al.
(Molecular Cloning:
A Laboratory Manual. 2r~d, ed., Cold Spring Harbor Laboratory, Cold Spring
Harbor
Laboratory Press, Cold Spring Harbor, NY,1989).
Host cells can contain more than one vector. Thus, different nucleotide
sequences can .
be introduced on different vectors of the same cell. Similarly, the nucleic
acid molecules can be
introduced either alone or with other nucleic acid molecules that are not
related to the nucleic
acid molecules such as those providing traps-acting factors for expression
vectors. When more
than one vector is introduced into a cell, the vectors can be introduced
independently; co-
introduced or joined to the nucleic acid molecule vector.
In the case of bacteriophage and viral vectors, these can be introduced into
cells as
packaged or. encapsulated virus by standard procedures for infection and
transduction. Viral
vectors can be replication-competent or replication-defective. In the case in
which viral
replication is defective, replication will occur in host cells providing
functions that complement
the defects.
Vectors generally include selectable markers that enable the selection of the
subpopulation of cells that contain the recombinant vector constructs. The
marker can be
contained in the same vector that contains the nucleic acid molecules
described herein or may
be on a separate vector. Markers include tetracycline or ampicillin-resistance
genes for
prokaryotic host cells and dihydrofolate reductase or neomycin resistance for
eukaryotic host
cells. However, any marker that provides selection for a phenotypic trait will
be effective.
While the mature proteins can be produced in bacteria, yeast, mammalian cells,
and
other cells under the control of the appropriate regulatory sequences, cell-
free transcription and
translation systems can also be used to produce these proteins using RNA
derived from the
DNA constructs described herein.
52
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Where secretion of the peptide is desired, which is di~cult to achieve with
multi-
transmembrane domain containing proteins such as transporters, appropriate
secretion signals
are incorporated into the vector. The signal sequence can be endogenous to the
peptides or
heterologous to these peptides.
Where the peptide is not secreted into the medium, which is typically the case
with
transporters, the protein can be isolated from the host cell by standard
disruption procedures,
including freeze thaw, sonication, mechanical disruption; use of lysing agents
and the like. The
peptide can then be recovered and purified by well-known purification methods
including
ammonium sulfate precipitation, acid extraction; anion or cationic exchange
chromatography,
phosphocellulose chromatography, hydrophobic-interaction chromatography,
affinity
chromatography, hydroxylapatite chromatography, lectin chromatography, or high
performance
liquid chromatography.
It is also understood that depending upon the host cell in recombinant
production of the ..
peptides described herein, the peptides can have various glycosylation
patterns, depending upon
the cell, or maybe non-glycosylated as when produced in bacteria. In addition,
the peptides
may include an initial modified methionine in some cases as a result of a host-
mediated process.
Uses of vectors and host cells
The recombinant host cells expressing the peptides described herein have a
variety of
uses. First, the cells are useful for producing a transporter protein or
peptide that can be further
purified to produce desired amounts of transporter protein or fragments. Thus,
host cells
containing expression vectors are useful for peptide production.
Host cells are also useful for conducting cell-based assays involving the
transporter
protein or transporter protein fragments, such as those described above as
well as other formats
known in the art. Thus, a recombinant host cell expressing a native
transporter protein is useful
for assaying compounds that stimulate or inhibit transporter protein function.
Host cells are also usefi~l for identifying transporter protein mutants in
which these
functions are affected. If the mutants naturally occur and give rise to a
pathology, host cells
containing the mutations are useful to assay compounds that have a desired
effect on the mutant
transporter protein (for example, stimulating or inhibiting function) which
may not be indicated
by their effect on the native transporter protein.
Genetically engineered host cells can be further used to produce non-human
transgenic
animals. A transgenic animal is preferably a mammal, for example a rodent,
such as a rat or
53
CA 02439401 2003-08-25
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mouse, in which one or more of the cells of the animal include a transgene. A
transgene is
exogenous DNA that is integrated into the genome of a cell from which a
transgenic animal
develops and which remains in the genome of the mature animal in one or more
cell types or
tissues of the txansgenic animal. These animals are useful for studying the
function of a
transporter protein and identifying and evaluating modulators of transporter
protein activity.
Other examples of transgenic animals include non-human primates, sheep, dogs,
cows, goats,
chickens and amphibians.
A transgenic animal can be produced by introducing nucleic acid into the male
pronuclei of a fertilized oocyte, e.g., by microinjection,-retroviral
infection, and allowing the
oocyte to develop in a pseudopregnant female foster animal. Any of the
transporter protein
nucleotide sequences can be introduced as a transgene into the genome of a non-
human animal,
such as a mouse.
Any of the regulatory or other sequences useful in expression vectors can form
part of
the transgenic sequence. This includes intronic sequences and polyadenylation
signals, if not
already included. A tissue-specific regulatory sequences) can be operably'
linked to the
transgene to direct expression of the transporter protein to particular cells.
Methods for generating transgenic animals via embryo manipulation and
microinjection, particularly animals such as mice, have become conventional in
the art and are
described, for example, in U.S. Patent Nos. 4,736,866 and 4,870,009, both by
Leder et al., U.S.
Patent No. 4,873,191 by Wagner et al. and in Hogan, B., Manipulating the Mouse
Embryo,
(Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986). Similar
methods are
used for production of other transgenic animals. A transgenic founder animal
can be identified
based upon the presence of the transgene in its genome and/or expression of
transgenic mRNA
in tissues or cells of the animals. A transgenic founder animal can then be
used to breed
additional animals carrying the transgene. Moreover, transgenic animals
carrying a transgene
can further be bred to other transgenic animals carrying other transgenes. A
transgenic animal
also includes animals in which the entire animal or tissues in the animal have
been produced
using the homologously recombinant host cells described herein.
Tn another embodiment, transgenic non-human animals can be produced which
contain
selected systems that allow for regulated expression of the transgene. One
example of such a
system is the crelloxP recombinase system of bacteriophage P1. For a
description of the
c~elZoxP recombinase system, see, e.g., Lakso et al. PNAS 89:6232-6236 (1992).
Another
example of a recombinase system is the FLP recombinase system of S cerevisiae
(O'Gorman et
54
CA 02439401 2003-08-25
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al. Science X51:1351-1355 (1991). If a cf~elloxP recombinase system is used to
regulate
expression of the transgene, animals containing transgenes encoding both the
Cre recombinase
and a selected protein is required. Such animals can be provided through the
construction of
"double" transgenic animals, e.g., by mating two transgenic animals, one
containing a transgene
encoding a selected protein and the other containing a transgene encoding a
recombinase.
Clones of the non-human transgenic animals described herein can also be
produced
according to the methods described in Wilmut, I. et al. Nature 35:810-813
(1997) and PCT
International Publication Nos. WO 97/07668 and WO 97/07669. In brief, a cell,
e.g., a somatic
cell, from the transgenic animal can be isolated and induced to exit the
growth cycle and enter
Go phase. The quiescent cell can then be fused, e.g., through the use. of
electrical pulses, to an
enucleated oocyte from an animal of the same species from which the quiescent
cell is isolated.
°The reconstructed oocyte is then cultured such that it develops to
morula or blastocyst and then
transferred to pseudopregnant female foster animal. The offspring born of this
female foster
animal will be a clone of the animal from which the celh e.g., the somatic
cell, is isolated.
Transgenic animals containing recombinant cells that express the peptides
described
herein are useful to conduct the assays described herein in an in vivo
context. Accordingly, the
various physiological factors that are present ih vivo and that could effect
ligand binding,
transporter protein activation, and signal transduction, may not be evident
from in vitro cell-free
or cell-based assays. Accordingly, it is useful to provide non-human
transgenic animals to
assay in vivo transporter protein function, including Iigand interaction, the
effect of specific
mutant transporter proteins on transporter protein function and ligand
interaction, and the effect
of chimeric transporter proteins. It is also possible to assess the effect of
null mutations, that is
mutations that substantially or completely eliminate one or more transporter
protein functions.
All publications and patents mentioned in the above specification are herein
incorporated by reference. Various modifications and variations of the
described method
and system of the invention will be apparent to those skilled in the art
without departing
from the scope and spirit of the invention. Although the invention has been
described in
connection with specific preferred embodiments, it should be understood that
the invention
as claimed should not be unduly limited to such specific embodiments. Indeed,
various
modifications of the above-described modes for carrying out the invention
which are obvious
to those skilled in the field of molecular biology or related fields are
intended to be within
the scope of the following claims.
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SEQUENCE LISTING
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<120> ISOLATED HUMAN TRANSPORTER PROTEINS,
NUCLEIC ACID MOLECULES ENCODING HUMAN TRANSPORTER PROTEINS,
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gcccctttct tcgtcctggg catggctctt atggcagacg gtgtagaggt gtttgtcgtt 540
ggcttcgtgt tacccagtgc tgagacagac ctctgcatcc caaattcagg atctggatgg 600
ctaggcagca tagtgtacct cgggatgatg gtgggggcgt tcttctgggg aggactggca 660
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ttcgggattg gaggagccat acccactgtg ttctcgtact ttgctgaagt cctggcccgg 840
gaaaagcggg gcgaacactt gagctggctc tgcatgttct ggatgatcgg tggcatctac 900
gcctctgcca tggcctgggc catcatcccg cactacgggt ggagcttcag catgggatcg 960
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gtggagagag ataaatatgc aaatttcact attaacttta caatggaaaa tcagattcat 1500
actggaatgg aatacgacaa tggcagattc ataggggcca agttcaaatc tgtaactttc 1560
aaagactctg tttttaagtc ctgcaccttt gaggatgtaa cttcagtgaa cacctacttc 1620
aagaactgca catttattga cactgttttt gacaacacag attttgagcc atataaattc 1680
attgacagtg aatttaaaaa ctgctcgttt tttcacaaca agacgggatg tcagattacc 1740
tttgatgatg actatagtgc ctactggatt tattttgtca actttctggg gacattggca 1800
gtattgccag ggaacattgt gtctgctctg ctgatggaca gaattgggcg cttaacaatg 1860
ctaggtggct ctatggtgct ttcggggatc agctgtttct tcctttggtt cggcaccagt 1920
gaatccatga tgataggcat gctgtgtctg tacaatggat tgaccatctc agcctggaac 1980
tctcttgacg tggtcactgt ggaactgtac cccacagacc ggagggcaac aggctttggc 2040
CA 02439401 2003-08-25
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ttcttaaatg cgctatgcaa ggcagcagcc gtcctgggaa acttaatatt tggctctctg 2100
gtcagcatca ccaaatcaat ccccatcctg ctggcttcta ctgtgctcgt gtgtggagga 2160
ctcgttgggc tgtgcctgcc tgacacacga acccaggttc tgatgtaata 2210
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1 5 10 15
Asp Ile Ala Arg Glu Val Lys Lys Gln Thr Val Lys Lys Val Asn Gln
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35 40 45
Phe Gln Asp Glu Glu Asp Asp Asp Asp Tyr Tyr Pro Ala Gly Glu Thr
50 55 60
Tyr Asn Gly Glu Ala Asn Asp Asp Glu Gly Ser Ser Glu Ala Thr G1u
65 70 75 80
Gly His Asp Glu Asp Asp Glu Ile Tyr Glu Gly Glu Tyr Gln Gly Ile
85 90 95
Pro Ser Met Asn Gln Ala Lys Asp Ser Ile Val Ser Val Gly Gln Pro
100 l05 110
Lys Gly Asp Glu Tyr Lys Asp Arg Arg Glu Leu Glu Ser Glu Arg Arg
115 120 125
Ala Asp Glu Glu Glu Leu Ala Gln Gln Tyr Glu Leu Ile Ile Gln Glu
130 135 140
Cys Gly His Gly Arg Phe Gln Trp Ala Pro Phe Phe Val Leu Gly Met
145 150 155 160
Ala Leu Met A1a Asp Gly Val Glu Val Phe Val Val Gly Phe Val Leu
165 170 175
Pro Ser Ala G1u Thr Asp Leu Cys Ile Pro Asn Ser Gly Ser Gly Trp
180 185 190
Leu Gly Ser Ile Val Tyr Leu Gly Met Met Val Gly Ala Phe Phe Trp
195 200 205
Gly Gly Leu Ala Asp Lys Val Gly Arg Lys Gln Ser Leu Leu Ile Cys
210 215 220
Met Ser Val Asn Gly Phe Phe Ala Phe Leu Ser Ser Phe Val G1n Gly
225 230 235 240
Tyr Gly Phe Phe Leu Phe Cys Arg Leu Leu Ser Gly Phe Gly Ile Gly
245 250 255
Gly Ala Ile Pro Thr Val Phe Ser Tyr Phe Ala Glu Val Leu Ala Arg
260 265 270
Glu Lys Arg Gly G1u His Leu Ser Trp Leu Cys Met Phe Trp Met Ile
275 280 285
Gly Gly Ile Tyr Ala Ser Ala Met Ala Trp Ala Ile Ile Pro His Tyr
290 295 300
Gly Trp Ser Phe Ser Met Gly Ser Ala Tyr Gln Phe His Ser Trp Arg
305 310 315 320
Val Phe Val Ile Val Cys Ala Leu Pro Cys Val Ser Ser Val Val Ala
325 330 335
Leu Thr Phe Met Pro Glu Ser Pro Arg Phe Leu Leu Glu Val Gly Lys
340 345 350
His Asp Glu Ala Trp Met Ile Leu Lys Leu Ile His Asp Thr Asn Met
355 360 365
Arg Ala Arg Gly Gln Pro Glu Lys Val Phe Thr Val Asn Lys Ile Lys
370 375 380
Thr Pro Lys Gln Tle Asp Glu Leu Ile Glu Ile Glu Ser Asp Thr Gly
385 390 395 400
2
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
Thr Trp Tyr Arg Arg Cys Phe Val Arg Ile Arg Ile Glu Leu Tyr Gly
405 410 415
Ile Trp Leu Thr Phe Met Arg Cys Phe Asn Tyr Pro Val Arg Asp Asn
420 425 430
Thr Ile Lys Leu Thr Ile Val Trp Phe Thr Leu Ser Phe Gly Tyr Tyr
435 440 445
Gly Leu Ser Val Trp Phe Pro Asp Val Ile Lys Pro Leu G1n Ser Asp
450 455 460
Glu Tyr Ala Leu Leu Thr Arg Asn Val Glu Arg Asp Lys Tyr Ala Asn
465 470 475 480
Phe Thr Ile Asn Phe Thr Met Glu Asn Gln Ile His Thr Gly Met Glu
485 490 495
Tyr Asp Asn Gly Arg Phe Ile Gly Ala Lys Phe Lys Ser Val Thr Phe
500 505 510
Lys Asp Ser Val Phe Lys Ser Cys Thr Phe Glu Asp Val Thr Ser Val
515 520 525
Asn Thr Tyr Phe Lys Asn Cys Thr Phe Ile Asp Thr Val Phe Asp Asn
530 535 540
Thr Asp Phe Glu Pro Tyr Lys Phe Ile Asp Ser Glu Phe Lys Asn Cys
545 550 555 560
Ser Phe Phe His Asn Lys Thr G1y Cys Gln I1e Thr Phe Asp Asp Asp
565 570 575
Tyr Ser Ala Tyr Trp Ile Tyr Phe Val Asn Phe Leu Gly Thr Leu Ala
580 585 590
Val Leu Pro Gly Asn Ile Val Ser Ala Leu Leu Met Asp Arg Ile Gly
595 600 605
Arg Leu Thr Met Leu Gly G1y Ser Met Val Leu Ser Gly I1e Ser Cys
610 615 620
Phe Phe Leu Trp Phe G2y Thr Ser Glu Ser Met Met Ile Gly Met Leu
625 630 635 640
Cys Leu Tyr Asn Gly Leu Thr Ile Ser A1a Trp Asn Ser Leu Asp Val
645 650 655
Val Thr Val Glu Leu Tyr Pro Thr Asp Arg Arg Ala Thr Gly Phe Gly
660 665 670
Phe Leu Asn Ala Leu Cys Lys Ala A1a Ala Val Leu Gly Asn Leu Ile
675 680 685
Phe Gly Ser Leu Val Ser Ile Thr Lys Ser Ile Pro Ile Leu Leu Ala
690 695 700
Ser Thr Val Leu Val Cys Gly Gly Leu Val Gly Leu Cys Leu Pro Asp
705 710 715 720
Thr Arg Thr Gln Val Leu Met
725
<210> 3
<211> 197997
<212> DNA
<213> Homo sapien
<220>
<221> misc_feature
<222> (1). .(197997)
<223> n = A,T,C or G
<400> 3
gcagaagctt tccattatga aaggcactgg ggtagtgctg tagggtcatt cagactctgc 60
tttcaactaa ccagaatttg gagctcactt aatggtattt ctggagctga ttatgtacag 120
gaagcagctg atatgcagtg tctgatgtgg tcatccgttc aatggatagg atggcagttt 180
attttattta ttgtgtgaaa tattcagatt ttctgttaat ttttaaagta ggaaatgtct 240
ctcaacttca tcattgtcct ctctgcccct tttccctgac ttcttgagtt tttgagcacc 300
3
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ccatcctcag cctccctttt caccaccttt ccattttttc ttactgttct catggatttt 360
cccatgtctc tcttcctccc tttctcttcc tctcagttct taaaaaaaaa aaaaaaaaaa 420
aaaaaaaaaa gagctggggg agaacaaaca gaaaaatcat ttactaaaac atttggttaa 480
aaaccttgca atcatatgtt tattttaatt tcattttatc attttccaaa acgcagcctg 540
tgagaaccat actcctgttt tgcctactgg gcatttgtag gtattcagtg ggaatgaagc 600
atggtccatt tataaacaaa catctgtttg tctttattgc tgggtttgtt tgtggcattt 660
ggaggagtaa ttaaatgtat gttagtaata gctactttaa atattttaaa ctagctacaa 720
aatgaaaaag tagaaatgac gattatataa atcaatgttt ggaatgctgc catagcttaa 780
ggcttgggga gttaatgatg ggggagaaag acttttttaa cctctgttat catctgtgta 840
caaggatgaa cctgtcagtg actcacatga gccaccagaa gaggcagctg agtggacaaa 900
tggaatataa gttaccttga ggtcagcctg aatttaagga accaaggcat gaggggacta 960
tcagtgaaaa agaaaaccaa ataacaggtc ctggaaggtc cagactttac taccaaaaca 1020
gcagctaacc agagtggcag gtgggaggat gtgagaggga gagagggaga gagagagagt 1080
gtgtgtgcgt actgggtatc cagagatgaa gaaaatagaa gtggggttgt gaagggacat 1140
ggcatggagc ttacctgtcc ctgaaggcct gggggtagtt tctgaaaggg cagactccca 1200
ccaaacaagc catatgttct aattcaatat caaatgcagc cataaaatca tttgatgaac 1260
tctgacaatc ctgtggatat gacactatgc tatgggaata tactccttgc cagtttgggg 1320
tgccatgtga gtctctgtta ccatttgttg gaaggaaaca caatgttaag taggggagaa 1380
attcaatttt acaaatatag ctatagtttg attgatagca attgtatatg gggctgcttg 1440
cttttcagtg tgaggaaata ggcccagtag gattcttcta atactctggt ttgtctttgc 1500
aatatttgta cgtgtctgtg agcaatacta tttttctcaa tcaaggtata tgcttgtagc 1560
atttcaaaat gtgttgggcg tgcttttaaa gatgcaagat gctctgtttt attctcaggg 1620
gaattcctgt gtactgttac cctggggata gaattcctta aaataaaaca ataataatag 1680
cagcgaaaac cctaaatact ccaagtgatg ctcacactta agcagagaag cagcctaggg 1740
tgtacatgta tatacagtga gatcaaaaaa aaaaaaaaaa ggaaaaacta attctgtgtt 1800
gtagtttgtg ggacactgaa aagaggtcaa gaaatagacg gtaaaaaata tatatataga 1860
aaggaataat aagtatctcc tctatttctt ctttcgcagt tctgttttga acccaaagtg 1920
gatgatgctg tcagagctga actactgaaa ggaggctgtg aaaatttccc atcttctcat 1980
tggccatcag ttgagataag atggaagact cttacaagga taggacttca ctgatgaagg 2040
gtgccaagga cattgccaga gaggtgaaga aacaaacagt aaagaaggtg aatcaagctg 2100
tggaccgagc ccaggatgaa tacacccaga ggtcctacag tcggttccaa gatgaagaag 2160
atgatgatga ctactacccg gctggagaaa cctataatgg tgaggccaac gatgacgaag 2220
gctcaagtga agccactgag gggcatgatg aagatgatga gatctatgag ggggagtatc 2280
agggcatccc cagtatgaac caagcgaagg acagcatcgt gtcagtgggg cagcccaagg 2340
gcgatgagta caaggaccga cgggagctgg aatcagaaag gagagctgac gaggaagagt 2400
tagcccagca gtatgagctg ataatccaag aatgcggtca tggtcgtttt cagtgggccc 2460
ttttcttcgt cctgggcatg gctcttatgg cagacggtgt agaggtgttt gtcgttggct 2520
tcgtgttacc cagtgctgag acagacctct gcatcccaaa ttcaggatct ggatggctag 2580
gtgagtgtgt ggtgtcagtg aggccaactc tgaaactggc ttatttgtta agcacagtta 2640
tcaacataga gaataaatac ttttcatcta gagtactgca ttttttctaa caaatttttt 2700
attacaaaaa actttaagca tacaaaagaa tcaaaataat tgtacagtga acacctgtgt 2760
atacccacca cctagattct gtagttaaca tttactatat tttctttatc atctctccat 2820
caacccaatt tatttttatg catttcaaag taagttagag tatgttttaa cttatatttt 2880
tatcatagtt gcattggttt ctcctttatt aaattccaaa gctactctct tggaagttaa 2940
taatgctgtt atatttaaca cttttctaaa tacagagatt agttctgtaa aactctgaca 3000
attgtgattt caaatgtcaa tttttatgtg taatacagta gtttaaatat tttaggaccc 3060
tgataacttt aattttttct attatctatg tgtatgtttg cttagatttt caactagcct 3120
tattcgtata attttctata ggtgattttt aatctttttc aagctgtgta actttgtgcc 3180
caaactagaa aaacttcatt tctttatatt attttaaata tataaatatt taaatgtgtg 3240
ttaaaaataa tataacctct ctataaaata tctatacagc agatttgatt tttgttttga 3300
agttaccatg gactcaggtt tatattttta caaagctata gttaacactc tccgagtttg 3360
tgcaagggaa aataagtagg taagaagcaa gcatgcagtg aactctcccc tcagcagttt 3420
aaggatccac agcaaacact tggtatataa aatcaggagg atttccaaac tgcttttgca 3480
aatcctgcct cagagaacac gtttatgtgc atgttaaatc tttatgtgtt tcagagtcta 3540
tgaaagaaag aatacttagt atctatgttt ccaagggact tctttaaaac taaagcttca 3600
aaaacaaaag tatacaattc ttgaaaacta gtaactttat taagtaatgt tattatcatt 3660
gaggctgaga aatttcctgg gtgattaaaa ttaaatgcta tatcctaatt tgttctgtga 3720
caacacaaga gagatactca tgaatgagaa aatgatatac atctttttat tcctttaaca 3780
cttgtttctt ttatatgacc cactcctagt atttgtttct ttatgcttag ctaaaataaa 3840
tggatccatg aaagtagatt gtgtaggttg aaggggtggg agtcaggagc atattgtgca 3900
4
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
tattttgctg gtacactcat ttgtgtgttc cactatcaaa tgcagacatc accacgcttc 3960
agaggagggt ccttggtcat ttgtcaaatt gatcatctca gcagactggt tttgtatcta 4020
gcactgtata tcatttttgt tttttttctt ttggtgagca agtatatcca actgtaagcc 4080
tagacatcaa aatatgtatt cagattttgc taaaggaata tgtcagcaga atgt.gtaaag 4140
aagatagacc ctgagaaata tactcacaca aatatttaat aattgccagc ttaaatttag 4200
aataaaaaat attttaatgt tatggtccca ccatttatac tggaaagact acagaaaaat 4260
ctaagccaaa cttgccctat gcctttatct tcctggtgct gtatgaaagc acagtatgac 4320
atacggttct cttatcaaac tatcctccaa acccagaaac cctggtgatg ttacccaccc 4380
ccaggtggca tttagaagtg tagataatgc tgcagaacaa ttggagcaca gatattctca 4440
tctgcccata aatgttgcct gctttagtga attaaaaata tgtccccagg agtgtatagt 4500
ttcactcagt gttgtgagtc cccaacagga gcattttaat gctttcataa ctgtgccttc 4560
cagacttgca tttcaattat tttgccattg taatttcact cataattgca ttctaacaga 4620
gtctttttag acaagcattt aaattgtttt gccattttca tttcatcgaa taatagcttt 4680
gtaatataat taccacacta agtcatagca ataggatcaa aatgcaagaa agcaggatgc 4740
atgacccatg tttggtggca ttttgaggat atagatttgc aaaataattt ctcatttgct 4800
actaagcatt gtgtcatttt cctcaaggaa gaccaggaac aaaaatatgg cccaatttat 4860
gttttaatta tttgggagat tgagcctctc tttgtgtttg ctaaaagatt gacttaaagg 4920
tatctaaggg catacagtgt atgcctgtat ctgtgtatat agtctgtgtg tatatataac 4980
attgttaatt gggccagaag aagttagagt aaatatttaa ttatctaatg atctgctctt 5040
tggaagaaca agggcagtca ctttaagata atctgttctc tattgccaaa gtctatttat 5100
taagcactta ctgtgtgtaa agcacagtat tgtcacaagg ataataggac aaaaaagaga 5160
ggcaaggttt ctctatccaa ggaagttata gtctagtaat ggagaaatca caaatgagta 5220
gattaccagt ctagatacca agtgtcagta gccataagac aagtgtaaat aaagagactt 5280
cgagcaggga gaacatgtta cagggtaagg ccctgactaa agcagtggag agaaggaatg 5340
gatttcagag ctactccaga ggtagagtca gcaaggattt gctcattgat tagcttcgag 5400
tggggaggag gggaccaatt aatgaaagaa tggcagttca ttaaattagg taaggcagaa 5460
tatagatttt agcaggatgg agaaagatga aaatgtttag tttgagaccc tctcattttg 5520
aggtccttgg ggacaatcaa agaggagtgc caatgggcta ttccccattt gagctgaaat 5580
gcccagagat ttggaagtca tcttgtttaa gacaacactg ccaatgacct taatgaggga 5640
gagaatggtt tcccgctgaa tactatttat ggggctggtg gtggaggagg aatcattaag 5700
gaggtagagg agaagccaga gaaaggagca gccaggagaa aggggaggtc tggaacttgc 5760
tggaagaagg aagggatgta tagctttcag gcgccgcagt cttgcagtag aggctggcaa 5820
ttaagagtta ttggctgctg cagacatctc ttaagatgaa cacagtgagc acaataaaca 5880
caatgagcca gactagtaag tttttaagac ttcttattga atgaagtgaa aatgtgtcct 5940
gctgatttcc agcccctggt cacaaggcag gcttttccag gacatcaggc attctccctt 6000
cccatttgct gtgactcaga gtggattctg tggaagtgag ccctctgact tagcttctgt 6060
tcagtggttg ttgagggggg caggtatggt tctaagatta cccctaggca ggaaagatgt 6120
gcaaccattt tcagctcagc cctaccccga ctctagtcca ttagggcaga acaagcagcg 6180
tgctggcacc tcttcccttg catcagaggg gctggcctcc cactcactgg cttatttcct 6240
cctaagaccc atccagtgaa gcagagaaaa tgatcattat tggttcaact gctttattgc 6300
tgagggttac aaagatgagt gatatttagg aactaaaata ttatcacagc caggtgttga 6360
ctaatacaaa atagatgcaa agcaacatat tctaacccca aatgtacttt tataactaaa 6420
cctgagctaa ttaatagtga aacatcctgt gcttctttta tccttttcaa ggcaaatatc 6480
aactgctacc cctaattccc gtatgcattg gtagttttca ggacaggagt tttagcagca 6540
taattgagac agggccagat tgctttggga agaaacatga atgttgttga gaaaaatgga 6600
gacagcaaga acatttcatt tgataagcat taattgagta ccaactctga gccaggtacc 6660
tggcaaagta cattgagagg atattctgat tagtatattg tgatctgtgt cattcaggaa 6720
cttttttgtt gttgttacaa tgtgacaaat acacttgtga taaattttag tgctttatgt 6780
tttttttttt ctgtgacaaa gatactgaat aaaatgctat aggattgcag aagagggaac 6840
taccaggctt acctttggaa actgatgaaa actcatctag gaattgacat ttaaacagag 6900
ttttgaacca tggcacgtgt atacctatgt aacaaacctt cacattctgc acatgtatcc 6960
cagaacttaa aataaaaaat ataaacaggg ttttgagagc tggacaagag tttaccgggc 7020
agataaaagg aaggatggca cttcgggaag atgttataat tgcccaatag cctggacttg 7080
gtaggcaatt atgtgaactt atctgtggta ggcgtagtag tgttggagct gacaggtcac 7140
attgtggata tattgtgaag accttaatgc caagataggg gtttggactt tatcatgaaa 7200
tcattagacc tgtgttttag aaagctgcct ctggcagcag tgtggagaga gaatgagatg 7260
ggagagagac ttaaaatgtc aaatagactt taagccactt acttgtgtga ccatttgcaa 7320
gtaatttaaa tttctctgtg cctcggtttt catagctgta cgttggcaat aagaataccc 7380
aattagattg ggaggattga atgagataat atatgtataa ctcttagcaa agcattgaat 7440
atatagcaaa cactaataaa tgatagctgc tataataatt aatagaaggg tgaatatcag 7500
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ttaggaggct aatgcaatag cccaggcaaa acatgaagaa atccctaact gtatcagtta 7560
atagggtgac agaggtgtca gaatcggaag catttctgga agagaaatgg aaaaatttgg 7620
tgaatgagaa agataagcag gtagttgagg atgatagcca aggctttggg caggtacagg 7680
gaaggcagag cagatgggga ggctgatgaa gttcattttg gagggtgtta gtctggtagg 7740
tagaagaggt caggaccaga atggcagatg tgagattctg aaaagggagg tttaaccaga 7800
gaacacagct aaacaggagg cggacaatga ctaatgaagg tatcagacag cttagcaagg 7860
ggttagagag tctttcggcc ccaaaataac taacagccag aagtaaattc ctctgcatga 7920
cacttttata ttttttaatt aaaaaaatca tattacaatc ctcttaaatc tcctgtttcc 7980
cttcagaacc agtagccatg tttaaaattt ggtctatgtg cttctctaga catgccaatt 8040
agatgtgctt gttctacaga gccctctttg aagctggagt gggagtgggg aggagggttg 8100
acaggacccc ttctggcatc cctttcctct tcttcatcag agcagactca gattcatcca 8160
tttaggtgag aaaggattcc atagctaaac aacaaaaaaa ttttaatatc ttatctggaa 8220
tatcagtgac ttcagctgtc acactagtgt tacaggttca gtgtttgatg aagtggtgtt 8280
atgaagtagc caggatgtgt gggtcatggc ccaaggccct gaaagagaga ttgagatgga 8340
taaggtttgg ctcctattac cagcgggaat cataatattt cccagctgtt gtttcctttg 8400
tgatttaggc aaaccctgtc attttaaaga tgaagatagg agacctagag agctcggttg 8460
gcttgcctga tctgtagctg ggtattagct gaggcagact ctggaacaca tataccaact 8520
cttagtccta tgttcctttc cagttccctc acgttttcat ttaaagcact ttcaatgggg 8580
ataaccctga gaagaacaca aggctctcca ttcttggtga aaatacacca ctctggtatg 8640
tcaagcaaga gaaagatgat ctggaaattc actttttgct tgtgaagtcc aacagagtaa 8700
tcacctgttg aacttttcaa aaaccttaat agaagatatc acctgttttt tgtaattata 8760
aacagtacat aatggcaaat atatggtcac aaatgagaaa tttgattgcc tcagagaatt 8820
aacctatcaa attagcccat agtgcaaagt catcttaaag aggcagagtg acagaaaaaa 8880
taaagaaagg gaatgtaagc ctcatgggaa ccagggattt tgtctgttgt tccctactgt 8940
atcctcctgc ccagatcagt gtgtggtaca taataggatc tcaacaaata tttgttgaat 9000
gaatgaatga cagtctctga gatctttaac actttgagaa ctaactcaag atatgaacat 9060
acctaaaata agaatgtact attttgggcc gggcacagcg gctcacgcct gtaatcccag 9120
cactttggga ggctgaggca ggcagatcac gaagtcagga gatttagacc atcctgacta 9180
acacagtgaa accccatctc tactaaaaat ccaaaaaaat tagtcgggtg tggtagcggg 9240
tgcctgtagt cccagctact cgggaggctg aggcaggaga atggcatgaa cccgggaggc 9300
agaggttgca gtgagctgag attgtgccac tgcactccag cctgggcgat ggagcgagac 9360
tccatctcaa aaaaaaaaaa gaatgtatta ttttgaattg aaactcagta gaggaggaat 9420
accaaagaga aagtcataga tgcagatgtt ggcagtgccc ctccaaatag tgctcagggt 9480
tgaggtgccc tgacctaaag atttagccta tctatctagg ggccacatgt atctctattg 9540
tagggaaaga tgcattagac atccttggcc tgggatttgt gaagacacga atgctgtagc 9600
taagcacctc tcattctcat tacatgttcc aaagcaactt aaggatttca aattcagctt 9660
tctctagagc ttgcccatca gaaatatgct ggccaaagat tgccagtttt cttgggagtc 9720
ctgtttttca ttttaattgg ccaactggca ttctgttttt ttctatcagt cgtggtgagg 9780
ttgaccattg ggagactgct tctaagagat agatctgtct ttagtgtgct gacatatcgg 9840
gtttctgaac atataagact gaggtccaaa ccagtcttgc atcctgggag acctacttgg 9900
gtgttcacct agtttcctgg agaaatattc caacttcata gatactaggt tttctttaaa 9960
acatctgcaa tattagctag agtcaattgg tatactggac gaaaacagag ctagaccaat 10020
aatatcaaag cattaaaaag acaatacttt tacatataac tggaatttta cttgactttt 10080
ttccattttt agaagctctt gaaagtattt tttttttttt tttttttttt gagacagagt 10140
ctcactctgt cgcccaggcc ggactgcgga ctgcagtggc gcaatctcgg ctcactgcaa 10200
gctccgtaga agctcttgaa agtttttaaa atatttctaa ataattctag aataaatata 10260
gatatttaca aatctgggat atatttttgg atagaaatgg agtagttgtt tttgttttac 10320
ttaaaacttt cctatcaact gtaccaagtc agactacttg gcattcctat atcttccatt 10380
gatagctaga gtgttcactc aaccaaaatt tattgagcat ctactatgtg acaagcacta 10440
tttggactct agaaatataa tagtaataaa acagaaaata tctctggtag atcttgagca 10500
tgttttcagc ttctctttct gtagttttta cttggatgct ggaattcctt gaaatataca 10560
gaattcattc ctcccatgcc cctttgcttt gtttgacctt ctaggaaggg tagtttcttt 10620
gagagaagag ctggatagat atgacctgaa acccttattc ctttagggat taacaaaagg 10680
ataaatattc tgttgggatt tctagctgct ttattgagag atagctgggt ctgaaaacaa 10740
cagcttcaag tttctaagat tcccctttgc tagtttcata gactcttatt ttccactaga 10800
ctgcaggtgg agtatagtga ctggctcacg gtgcattgaa ccaatctttt tagagtgtgt 10860
ctaacccctg ctcagtgctg ccaggtagac cagcatatcc aagctgatga catagaggga 10920
cacagagttt ttcctctttt agctttattc caaatttgtg atgatttggc atctgagtgt 10980
ggcctaaatt ttaagaccca taactttttt tccctacaag tgtacactgc agactttagg 11040
atcagcagct ctataccctg cttgatctct ttgtcctcag catctatgtg ggtacctctc 11100
6
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
atggaacttt attctttgtt gtgtatgcac cctctgtctc tgtttagcct ctcaggcatc 11160
acagtgatcc taattggagt taaaaagtta ttggctcata gagcacatcc tgtctaccaa 11220
cactagcaaa atcttatttg gattcattat tgcaaaattg gaaaagaaga ttcaataatt 11280
tatcgccatt atggtcctgg atcttatttc ctcttgcctt tcaaattctc ctgggaggag 11340
agaggcttct gacatccgag tggaggtgga ctgagcagcc tgaaaagact cttccatcca 11400
aagagtcggg ttcactttca caggacagga aatgaacaga aactctctgc ctcattcctg 11460
aaacttgcca ataagtagca ttctaagacg gtgagaattt gataggaaaa tatccttctg 11520
ccaccctgaa ggtcactgag ggttttccat aatacagctc tgaacctgaa ggcagatttc 11580
tctatttcta cactctgaat cattgtatcc ctatatccta gaacactcta ggatgaggat 11640
tctgatagga atcctcacta acgctgagag cactggcctt acatacattg atccagtctg 11700
agactctacc taaagtcact ggagaacttg ccgatgtatt tcaggttgga gaatttgttc 11760
atgtggactt cactttaaat aaggcaattg aagccaggcg cggtggctca cacctgtaat 11820
cccagccctt tgggaggcca aggcaggtgg atcacttgag gtcaggagct caagaccagc 11880
ctggccaaca tggtgaaacc ccatctctac taaaaataca aaaattagcc aggcgtggtg 11940
gtatgtgcct gtagtcccag ctactcagga ggctgaggca ggagaatggc ttgaacctgg 12000
gaggtggagg ctgcagtgag ccaagatcac gccactgcac tccagcctgg gtgacagagc 12060
aaaagtccat ctcaaaaaaa aaaaaaaaaa aaagtaaaat ttaaaatatt aaaaaattaa 12120
aaataaataa ggcagttgaa ttgttaatta tctaacaatt ggatcaagcc aaatataata 12180
atacagactc ataggtttga tttcagatta tcagtatgca gttgcaccct ttcatgtgaa 12240
atctaacatc caagtattct cacataaaca agagcaaggg cctttaaatg gcctgagatt 12300
actgctgaac acttcagtgt aagaaactgt tcatcagatt catcttaatg aattagtctg 12360
ataaattact gattaagtat ttctaacaca catatttgca aaccaacatt tttaccctaa 12420
atttagaaat ttctttttgg cctgtaagat tgtgagagaa aaagtaaaag aagtaaatga 12480
gtgtagttct tacctttaat ttcaaaattg aggtctctaa tcaaattgaa aatgatgagg 12540
aaacaatgag tttttccaat taaaaatttt tttactgtaa ggaatttgct taaaaataga 12600
atttaatgat ctttaaatta ttttcacatc tctctgtctt gattctgttt aatgtgctta 12660
taatcataac tgcaattatg taaatacaga ttagttcatc tttgctaact tcacattatt 12720
tcacataaag attttggaga atcaatacag tgtgttgtgg gcatcagtga gtatatggtc 12780
attcataatt cactgttcca gtcccttgct tagccatctg ttactgagca tttgaatgta 12840
tccagttctt tgcttattat atgaaaataa tgctccaatt aatgtcatta ctaaaattac 12900
tcttttgttt gggacattta gttggcatgt tttccccaaa gcaggagtga aagttactaa 12960
tgagtatatg atattttaat ttctttttta aaaacattaa cataagtaat actgaaagaa 13020
gaaaataata tggcaaatat tcaataattt tggaaatgcc atacaacaca ggcccaggga 13080
acagtcaagg ctgactagga agtcctttgc atattctcca aatatggtat ctcttcaact 13140
ttttcttttc cttttttttt tttttttttt ttttttgaga cggagtcttg ctctgttgcc 13200
aggctagagt gcagtggtgc aatctcggct cactactgca acctctgact ccctggttca 13260
agcaattctc ctgcctcagc ctcccaagta gctggggtta caggcacctg ccaccacacc 13320
cggctaattt ttgtattttt agtagaatgg gatttcacca tgttggccag gatggtctcg 13380
atatcctgac ctcatgatcc acctgccttg gcctcccaaa gtgctgggat,tacaggtgtg 13440
agccaccgcg cctggctggc ctcttcaatt ttttcttaag gaaattggtt gcaatgatga 13500
tgatgaaaag gcattagtgt gcccagggga aaacattagt ggtcagatga tgagacacag 13560
agagacgata aaggtgtccc atcttaggta cttaccctcc aaacactgaa gtagccccta 13620
atcccttacc ccagaaacga gttcattctc ccacgtccaa atctatgtta gcatttctca 13680
accggagccc tagtaacagg gacaggtctg tgctgtgtga cactgtctca cacattgcag 13740
gatgtttact agctgtggcc tctgtccaca aaatgtgagc agtgcttccc aggcattcct 13800
taaaacatgt gccttaccac actccagaca aggcctctca aatacatatt tctacctgac 13860
ccccaggaag caataggatc caggactcag gctctggtgg tggtattacc acaacagcag 13920
tggagactgg agttcagtgt gggggaaaca gaagagttgt ctcttgtgct ggctttctgc 13980
cttcctagta gtggttctac atcttctagg attgtggttt taacatcgag gatcttctac 14040
tgatcttgac aatggtcttg aattttttca aactaacgaa tatgtattag aaaatcaagc 14100
tggtcatcaa ctgagccgtt cttggagttg tagagcttag agaggaaagg tgatttagtg 14160
ctcacagaat caaaaaccct aaatttgaag aagagaaaaa ctgaagtcaa ataaagagga 14220
aatgattcac atagaggcac atgccattac cagtgaggta ggaataaaaa cttgatggcc 14280
taattttcag actgggattt tcttcatcca aaaagtgcac aatctcagtt ggtggtggtg 14340
gttacgcaac aatgtgaatg tactcaacat cacatcatag taagatgata aattttatgt 14400
tctgtgtgtt tgatacaatt taaaaaatgc agtgttccta tgcatttagg aatgaagagc 14460
tatttatcaa tattggaata ttgcttattt ctgcaataaa atttattttt tcactgtaaa 14520
gaccgagcct ggaaacagtg aagctgacag ctgagaaact gctgaaaacc ctaacgaaag 14580
cactttccag atccaactta ccttgtgggg ctgccatgta aaattgtgca ggttgtgacc 14640
tgcacaaaag gacctgggtt tgtggaaggg gtggggttgg aatccagccc tttcactcac 14700
7
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
cattcatctt ggtttagggc tgcttttatt tatctggagg aaaggatgat ctttgctaat 14760
tcctcttcct tgaaggagct cctttacgta attcatccac ccagatgaat ctgccttttc 14820
ccaattcatg tagaagtgtc ttctgtgcca ggaggctctg ttacttgatg atcaacttta 14880
gacagtgggg aacctgagga gatacaaaag gagaagggat ataagaaacc atgaccttac 14940
ccttccaaga actgagggca gatgcagaat attcagaaga tgagaacctt ttaaagggaa 15000
aatgtaacta aagtgcaaat ataattcagt aaatattcat gtaagcagtt acagttctgg 15060
attttaagta cagtcagatt gataggaggt ttttttttgt tttttgtttt tttagcaaat 15120
catgtgtcta attaacttaa aatatttggg aaattctgag aaatgaggga attttgagtt 15180
tataagtaca ctgcatgtta atgacccaga ataatagaat tggagagaat ggaccatctt 15240
ctctagctga ggcccaggcc caagagaaga gctcacacag agcagaacta agctacagcc 15300
accagggatt ttgtggggaa gattacctcc atactgggag gggcccatgc aatttgcatc 15360
accatagctg ccctctccag gagttcttgg gagagcaatt ctgtttaggt acatagcaga 15420
agagcaagct cctagagtgg aagttaaagg ggctgcttaa tgtttggtgt gtactttgtg 15480
aggttaggga ggtaagatgt taaagtcaga tgctgctcta cttagtatgg gatgtcaata 15540
aggtacttac ttcaaagtag gtactggaac atacctttct tctaagagac taagattttg 15600
ttgttgttct tcagagaagc atttgtggaa tatttctgta gtgcttgaat ggccaaagat 15660
caataaacaa aataaattac agtcctggtg gattttaata tttaaattta gataattatg 15720
atgtccaaag agaagagaaa gagatgcttt aagtcattac caaaaatatc ttagtccttg 15780
agagggattt gctgaaccaa agtgagaatg aagtctgtgt gtcccctttt ctgcaaagaa 15840
gctgacgcca catgatgcaa aggcctgccc ttaacccagt taatatgaac atggcatggc 15900
accaaagagg agacagaatc gacagggaga gtcccagttt taagctccca agcaagatct 15960
ttgagtgggg atgggagggg tccttttacc ctttcctaat gactaaatta aaccaaatct 16020
acccaagtgg cccctcctct cctcctggga gatggtggtg ccattgtcaa gataggtggg 16080
acacatttgg gagagattag gggaacacag tttcaccttc acatttaaga atccaattta 16140
ctcctgtggc cttgtttgaa ctctccttcc aacactcata taataataat tatatgtcct 16200
tgccttctgt aagctcattt cagttggtgt agcaaatata tttctaccac acacacaaaa 16260
attcttctct taggggttca tgtggaaatg aaaacacaat gtgttccact ccacgtgaaa 16320
agcaaggctg ctgtctctgc agcaaagtgc ttcctgacaa ggactgagct gaagtgtcat 16380
gaagctactc acctggaggg ggcatcagca tctcctgcct cttaagtcga ccagggaaag 16440
cttggacaag agcagccatg taggtgttac aggaaaggtc ctgatccaga cccctagaga 16500
acgttcttgg atgtcgcgca agaaagaatt cagggtgact ccacggtaaa agcaagttta 16560
ttaagaaaga aaaggaataa aagaatagct actccatgga ccagccccga gggctgctgg 16620
ttgcccattt ttatggttat ttcttggtga tatgctaaac aagggatgga ttactcatgc 16680
ctcccctttc tagaccatat agggaaactt cctgacattg ccatggcctt tgtaaactgt 16740
cagggtgctg gtgggagtgt agcagtgagg atgacctgag gtcactctcg tcaccatctt 16800
ggttttggtg gcttttggcc agcttcttta ctgcaacctg ttttatcagc aaggtcttta 16860
tgacctgtat tttgtacaaa cctcctatct tatcctgtta cttagaatgc cttaaccttc 16920
tgggaatgca gcccagtagg tctcagcctc attttaccca gctcctattc aagaccgagt 16980
tgctctggtt caaatgtctc tgacatgggg gctaatgaca tcagaactgg aaagaaagat 17040
gataaaggaa tgagtttttt taaaaaaaaa aaaaaaaaaa aaaaaaaacc aaacaaaaca 17100
aggataccaa atggaaaata aacaaaaaaa aagttttaaa aagaaaaaaa ttaagtgctg 17160
actgtgctac aacatgcatg aacctgaaaa acatgatatt atgccacaca caaagggaca 17220
aatattgcct atgaattatc tagaataggc aaactcatgg agatggggaa tggattagaa 17280
ttatgaggga ctcttttaga attatccaga gcaccatcca agggagaggg gaatgggaag 17340
gtacaatgtg tacagctagc tataggtttg tggtgatgac gaagttttgg acataggcaa 17400
tggtgatggc tgacagcatt gtgaatgtaa tcaataccat tgaattgtac actcaaaata 17460
attaaaatgg aaaattttct gctacataaa ttttaacaca atacaaaaag aaaaagtcag 17520
aaagaagtcc ttttggaggt gagtgggtgc tcctagtcac actcagagat agggagggct 17580
cactcttcct gtgtttgagg agatcctggg actggcagta catcctgtgt gtatatgtag 17640
aggcagatat gtcctaacag gacagccttt tctactctgt gtcttgaagt ttcacaggaa 17700
tcaggatgca tatgtggtcc caattctgtg tctgtgccag ctgttcctgt agctttcttc 17760
ttgaggggtc cagagcactt cagcatggag tttgccacaa cggcctctcc tctgtctgga 17820
ttgctgaatt cctgttgagc ttcctgggga atttgagagt agagggtggg acttggctat 17880
ggccgaatta ggctacttgg aaatcggagg ggaacaagta tgcaacctaa aacattctgt 17940
aggtcatttg tagttcactt gcagtcagat aaaatcatca ttagattaaa aaatgtgact 18000
tcatttatgt aacttttttg acaagatatt tattttaggt aaataaagct tacctctaca 18060
agagtcaaag ctctttttgc ttaagcttat tttttaaatg gaaatcaaac atgttgtata 18120
tacccttttt gaaatgtatc atacataatt taaacgcttc ctgatgattc acggtatcta 18180
tttttatttt ttatgtaata catttttttc ctatacacct ttctgtcatg ctgttattgc 18240
tgtcatttta actgccccta attcattctt tgatgtttct gtgctatggt ttaggatgct 18300
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
aaataaagag actcattaag aattatatat taaataagag caaatagtta tggagtcatt 18360
gcttggtttt acaaccgggc tttgggccag atatgtgggg tgtccagaaa ccaagttcaa 18420
ggttctggaa gagtgggcta aggacactgg aaatttggct tttgatatgt cactatatat 18480
cttttttttc aaaccgcaaa tagctctctg caaacagggc cagtgctcaa ctgtgcacca 18540
gttagaaaag tcaagtagta ggcttaaatt atttttatgt gtctggccag ggtataaata 18600
gtttaagagt ttagaattaa tacaaaatgg atttaagatt taaaattttt aactccctta 18660
ttctttgtta tatggtcact ggccgtgact attcttttca tcctctgggc ttcctttttc 18720
tctctctctc tttagagaca gtgtttcact gtgttgccca agctggagta cagtgatgca 18780
gtcgctgctc actgtagcct caacctcctg ggctcaagta attctcccac ctctgactcc 18840
tgagtatctg ggactatagg tgcacaccat cacatctggc taatttttta aaaaaatttt 18900
tgtagagtca gagtctcact atcttgccca ggctggtctt gaactcctgg gctcaaatga 18960
tcctcctacc ttggcttctc aagtgctgag attacaggcc tgagccacga tgtctggcct 19020
ctggcctccc tttctaatgg gtcttgtgat agaggctttt gtccccacag cctacttgcc 19080
attgttgttc tgactattcc tgggagaaag tgggtggagg gttcaggtca catctagtct 19140
tggctagtta gtacatagat tgttttaaac gtctttgaat ggccttcaaa actgcacaat 19200
ttgagcctgt ttttccagtc ttaatctgcc actcactctc ctccccaaga tattccaagg 19260
tcggggtagt tagggttgcc agatgcttat attaaaaaat tatttgttgt tgatctgaaa 19320
ttcaaatgtc accgggcaac cctaccacta gtgttgatga ccacaggagg gaggagagac 19380
tagagtttga ttcttgcctt ctgtatgcca ggcatggtca tatcagtcat attaatttca 19440
ttatctcatt taatcagaat cacatccaat cccagacctt ctgaatcagt gctaggtgtg 19500
gagcctggga aattgtgtgt atgtcttatg agcattccaa gtgattccta ttcaaaggga 19560
gttacagaaa caccatgacc tagaccagtg ccccccagac atcatgcaga tgcaggtcac 19620
ctgaggttag agctgaagtg caggtttcga tactgcagac ctggagtggg gcctcaggtt 19680
ctgcatgcca acaacctccc acgagacgct gaggctgctg gttgtgaatc ccacttggag 19740
tggtgctgtc ctgagcctcc cagccaagaa gcagcagcac ccccagcagt tgtgataatg 19800
aagattataa atgatactat gtttttaaat aatcagaggt ggggatataa gtggaagtat 19860
ttggcttatt caccaatata cagtaacatc tatgtatgaa gggtgaagtt ttgtgtgcca 19920
ttttttagga aacttttttt tttccaaaaa gtaaatgtct tacccgcagc tgagggatgc 19980
acattttttt cctccactaa gtggatattg gcttatttat gctggtcttt ctttcttata 20040
cacagtgggg ggacatctgt ccctaatcac atttacagct tcaaatccca gaatggactc 20100
cagatgtgca gggcacagtg tgacttggct ttatggaaat agtttctcag tgccagagtc 20160
atggtgatag agttataatg tgactccttc cattttcttc cctcctagct gcatgaaatt 20220
aaatggtttt attgagagta gcttagttta ctcagcagaa tctagttaga tcagaaatat 20280
ttaaggtaca actgtgtcaa agaactgatg gaaaatcgaa agagttaatc aaactgctca 20340
caaggacaaa agaaagtcct gctgcacgat cactttagcc ataaacatgc acactcaaat 20400
gaaggcagat tatgtaggtg acaggtggtc agcttctcag tccctttatc tgttgattta 20460
atatcttttt tttttttttt tgagacagac tcttgctctg tctgcaggct ggtgacaacc 20520
tgcaacctct gcctctcggg ttcaagtgat cctcctgcct cagcctccca agtagctgag 20580
actacaggtg tgtgccacca cgcgcagcta atttttgtat ttttagtaga gatggggttt 20640
caccatgttg gccaggatgg tctcaatctc tcgacctcat gatccgccca ccttggcctc 20700
ccaaaatgct gggattacag gcgcgagcca cggcacccgg cctctgttta tttttctaat 20760
ttgtagttat gagatgagca gaggttagat tggagatgtc tttgtggaat atcacctttc 20820
aaaatttagc agttctcaca cagaaagcac accagcagtg gcccaactat tagtctgttt 20880
attcatcaaa ttcttttttt tttttttttt tttttttttt tttgagacca ggtgtcactc 20940
tgttgcccag gctggagtgc agtggcacaa tcatggctca ctgcagtttc aacctcctgg 21000
gctcaagtga tcctcccacc tcagcctcct gagtagctgg gaccacagat gtgcactaac 21060
acgcttggct aattttttta tttttttgtg gagatggggg cctctctttg ttgtccatgt 21120
tggtcttgaa ctccgagcct caagtgatcc tcccaccttg gcctcacaaa gtgttggcat 21180
tacaggcttg aaacatggca ctcagcccat tcaccagatt cttacctagt cggtgcaagc 21240
cactgtaatc tgttgtatgg caaatacaaa gatcaaccag atacaaacta tctgcaaaag 21300
tcacttgtca tcagaatgac acaggaagca aaggactgag agattcattc tgtttgaagg 21360
aggagtggat ttttaaaaaa ggctttcagg agactgtggt atttgagcca ggcttcaagt 21420
tgtggataga atttggactt gcagagatgg agctgggtaa atggtgtttc tgaaagacag 21480
aacaaactga gcaaaggcac aaaggcatgg aaatcttgga tggagtattg tgaatagatt 21540
acagttccag tttgtcgaag tgtagcatgc ttacagggaa gcaattggaa acgtgggtgg 21600
gacctacctg ttgttggaga gcttgaaatg aattcatttt ggtgccaatg gggagtggtt 21660
gacgtgtgag cagcagaaga gtgacatggt gaaatgtcat ttgggagact gctgtgtgtg 21720
gaatggattg gacaggcaca aacttgagat gggaatcagc tgagacacat ctgaaatagt 21780
ctagccttca ggtgatgagg gccttgttgt gagtgatgga ggggagaagg tgtttatggg 21840
caaattcctg aaccttaaca acCatgagct atgggtgcag ggaggagggc catgtcatat 21900
9
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ggtactggga cctccagacc aggcgttatg ccttaatacg agagttctga ggttggcggg 21960
gaggtgggag agcccacttg taggaaggaa gatgtgtttg gttatcctga actgtaacag 22020
caagaattga gtgtgtcttc ttcacacagt gcccttagca gagcctgggt ggcaagtggt 22080
gccacccagt gaatctttgt tgaggttgag atgccagcca agctgctaag aggaagctgt 22140
ccaggaaggc ggttggaaat ggctcctggg gcctcacggg gaagctagcc ctagagatgg 22200
aacttgggca atcttggaat agtgtaacag agccctcagg aaagagaaag ctacagacgg 22260
agctttagtg atgccttagg gggtgagaga agtcctaaaa taaatggttt gaatgtaggg 22320
aaagtctcag ttgcctggag gccaagaaca aaaaggattt caggaaggag caagtgggca 22380
caatagatga gtgcttctga ggggacgggg ctgagaaaag gacacaggca gtggccatga 22440
gaaagctgtt ctggagggtg gcagggacag atgtcagatg aactgctcga tttggtttta 22500
atgtagattt ttaaagggaa ggaaacaagt cagtgtttaa acattggtta caaggtactt 22560
tgatatattt tttagataac gcttctaata acaaaagtat actatagact agagcagctg 22620
aaaaaatatt agtcatattc tgcaagggaa aatttccatc tttgtagcca tttgggtcat 22680
taagtgccag taatcacatc cagggtatgt atatatagga gatacgttgg caaaatagga 22740
gaggactggg tcagctgctg ctatctcatt ttatttattt ttccaaattg aaaactgtgt 22800
cagttacaag tacctctgga ttaaaaataa aatgaacttt gctgaaatgt taacatgtgg 22860
atagtcaaat agcaattata atgtggtaca gaattgccca gggcagaggt gtggtgggaa 22920
gacagctacc agctgatcac tccttccttt gatttgagct gactaaagtc agaggatcct 22980
ctaacactgc acactacaac tttgtccctg tgggtttctg ctggatcagt aaagaattct 23040
tctgcccccc agcccctccc aacactgggc acactaccgt tataaggtag ggatgtaggc 23100
agttatgtgg ctgatattaa tgtgatgttt atccctgtag gctggtactg cacatcacat 23160
ggtgtgtggg atcacagatc atattttttc tgccaaaagg cagtgaatta ttcccccaga 23220
gatgccatta tttcctcctt aaagagaaat aaattatata gctgctatat gggagtggca 23280
caccctgggt ggatcactct gaagcctgaa gcccaaggga gaacccagga aaggccaggg 23340
tactgccaaa ttctctccag cctgggaacg ttgcttcctc caaaccctct ggggcatctt 23400
gagctgctcc tattttgtag ggaccccgag ctctttcccc agctcagagg ctgaaaggag 23460
ttcctaggag tagcctctct ccctgagctt cctctcatct tagtggactg acttgagttt 23520
ggagggaact aagttgggag gctgagctca gtaaaatgtt gaccacccca cccttcctga 23580
tattgtacct tccttcggcc ttggcagctc agcatccttc tgatgtctct cggctctcct 23640
aatgttcctc ctgtagtctt ctctctgctt cctgaagagg gacttcccaa gagttggccc 23700
ttggtgcttg gttgtctgtg ttttgtgact caaattgcct ttcttagtca tccagcaaac 23760
acttatttca tatctacctc tgctacaggt ttcagactgg gtgatggagt tgcaaagcca 23820
tgatccctgc tcctgagagt ttacaggcca gagagggaca cagacatata acaaagtagg 23880
gcacctaatt tttacagatg ctataccaag gagattcaaa agaaggaatg gttagtgctc 23940
tcagagggca agggcaagac tggaaagatt tagtagaggc aaccttagac ttttctgatg 24000
atttccaggc ttgtggcttc aaatgttttg ttagatattc ccattaggtt gtgcaggaaa 24060
gaaagaaaga atggacagag gcttggaggc agagctggac ttgaatccta gatctgtcag 24120
ttaggagttt accactctgg aacaaggaaa caggtaatat tctttatctt gcagattgaa 24180
aaatactgtg atcatcactt ttccagagct tctcatcctc agttcctaat caaggggtgt 24240
tagcatgcag acaagaagca gggtaaggag actcatcagt gacctgttag tagtcatgga 24300
ctacaaggta aagaaagcat ggaccacaga gtctgttacc ttagtgaggt gctgtcagga 24360
ttggtgacaa tgtacaaaaa gcatctagta ctagaacagt actaatcaat acatctccca 24420
ctcattgtca acctgctacc agaaagccaa gtgactcacc ttccaaaaat gccctgttcc 24480
ctcctttttc ttccacagct gtcagtgcta atggtgtcct ccaggatcac tcagctttag 24540
tatctgcctt ttctgtcttt tgcttccttc gtattccctt gccttctctt tctgttaatg 24600
tataaaacca gtgatggcct gatttgaatg taagggaaga tgatgaccac tggggtcact 24660
gagagccaca gatttaagag ctcttaagca ttcattgttt gaaacaggat ttccctagag 24720
gaaatttgtg catttattca atgaggatgg aatcactagg tggtattcag catctgtctc 24780
agagctcatc ctgagtgcta cgctgcatea cactccggac cacacagagc tgcatgcata 24840
gcagaatttg tactctgcag actctacgga cacctggaac ccttaagagt agatgtgaat 24900
gtttcacttc cgccatgata gcatgagttc tgtggacctg atccacagaa aaactaaaaa 24960
ttactaaaga acataaaact aaccacttaa agtatctgga aatggcctgt aaaccaaaaa 25020
gtatctaaga caggtcttaa tcaattttga ggtttatttg ccaaggttaa ggatgcgtcc 25080
agagaaaagg aacacaaaac cacaggaaca atctgtgata catgctattt tccaaagagc 25140
gttttgggac ttctatattt aaagggaaaa agagcaggta gtagcacaaa gaggaaagag 25200
aaaaataaaa gaggagggta gataaaagaa gagaggggtt gcattccttt gagtctttga 25260
tcagcattca ctgaattcac attttacctt tgaaaagaaa gcatagagaa atagtcatta 25320
tgcattcttc tcacactcag taaattagga ctctacctaa gataaagtaa acatagagtg 25380
gctatttgtg gagccatctg gccttctatc tagccttctg tcttctaagt tacttgttta 25440
gaaacaaaaa gaaaggcagt tgcttacatg actcagtttc cagcttaact tttccctggc 25500
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
atagtgaatt tgggatgccg agattttatt ttcctttcac caggcatatg ggcattcagc 25560
aaattaagaa atacttattc aggaaaatct actaaaactc agttacaaag gtgagagctg 25620
tagtatttga agcaagacct ccacttttct tcccccactc caaactcaga gagatggaaa 25680
cttcactcta gactggtgca accaagaatg cagagctcct tctaccccag ctcctaattg 25740
gagacctgtt gtcttggaaa ggacagtatg tcagcctttc tcatcttgcc ctcagcaacc 25800
tgtttttgag actaagttcc aggcaagtgc agcccagagg tgtggggttc tttctttcat 25860
ccagcctcca cccataggac agaggaatct accttgaatg tggcgtgctg agaatcctgg 25920
gacctacatt atccttgccc ttgcttgaaa gctgtggttt ctcacaggaa gaagcaagct 25980
gagaagacct atggttactg ccccaccacc actgagaact cagctataag agtaggggtg 26040
tcacttagag agaagcatac cattgtctcc atctgcagtt ctagagccct aactcagaga 26100
tttgcctggg ggagaagtag gttgtaaaac agagacctct cagtatctta ccaaaggagc 26160
tgactttgtt tgcaacaaag tgtggagaaa tttatgccca aggtttctct caaaaaaaaa 26220
aaaaaaaaaa aaaagtagag gttgtagtga aaggcaatgg ggaagagatt ggtagattca 26280
ttggagatat agattaagtt gtaggccagc tagtttgctg ggaactaggg aaagagacag 26340
ctgagaggag cattcctgga gtcagagtga atcttaagca ctggccttgg aaactgtctt 26400
ttcaaaggag ccaaatttga ttgaatcaat ctaggaaaca acatataccc atagcaatca 26460
accttcagtt aattcaacag ctaggaatgg tcagggaaag agatagtgaa acagaatctt 26520
gccaaagccc tattagggta gctgtggaca tacccaaggc tacatcccct gagtggtaac 26580
atcagaggct tcattctatg gggggaagga aacagacttc agtaaaataa tccaactaat 26640
cactaaagaa ataaacaaat cacaatagca atccctggag agagg.gagga tctatcatcc 26700
tgattgctac aatatgttat gtagaatgtc cagtttccaa cagaaaaact atgagatatg 26760
ccaaaaaacg gaaagatatg acctatacac tagggttagg ggttaggggg cattaggcaa 26820
tagatactgg ctgtgagagc aaacaggtgt tgaatttaaa agaaaaagac atcaaagtat 26880
gcataataaa tacattcaaa gagctaaagg aaacccatga ttaaatacat aaaagtaggt 26940
attatgacaa tgtcatatca cacagagaat ataaatgaga tataaatttt aaaaaagaac 27000
caaatgaaca ttttggaatt gtaaagtata ataactgaaa tgaaaaatta accaaagagg 27060
ttcaagagta gatttgaact gacaaaagaa agaattaatg aactagaaga aagattgaca 27120
gaggttatac aagctgaaga gagaaaaaaa tgaacagagc cttagagaat tacaagacac 27180
cattgagtgc accaacctat atgtaatggg aatactggaa ggagaggaaa aagagaaagg 27240
agcagaaaaa atattagaag aaataatagc ttttcattat ttccaggctt tccaaatgta 27300
ttggaaaaat attaatgtac atatctagaa agctcaaaaa actccaagga agattagcaa 27360
acacaaagag gtcagagaca ttatagtaaa aatgctgaga atccaaagac aaggagagaa 27420
tcttgaaagc agcaagagaa aaatgactta ctacttataa gggaacccaa agactaacct 27480
gacttctcct cagaaatagt agaggccaga aaatagtgat gtccagattc aaattacata 27540
aagaaataaa tgttaactaa aaatcttata ttcagcaaag ctatctttca taataaaagc 27600
aaaataatga tatccccaga tgaacaaaaa cagaaataat ttgtttctaa cagatctgcc 27660
ttacaagaaa tgctaaagtt atttagtctg aaagcaagta acttcacaca ataatctgaa 27720
tcttcaagaa aaaaacaatg agcatatata tgtgtaaaag acagtattaa tggatatatc 27780
acctcccttt ttaaaaactg atttaaaaag caattgcatt aaactataca catatataca 27840
cacatacata tatatattat cattgagact tcacttatag aaatgtagca tatttgctaa 27900
taatagcaca aaggaggtag gtggaggcaa tattatattg ggctaagaaa atgactctag 27960
tggtaacgaa ttcacataaa caaacgaagg gatccacaaa tgataaataa agttaacata 28020
acaaaagata taaatatgta tatatatatt tgttctcctt tcttcactta gcttccttaa 28080
aagacata.at gttatataat gtaataatta ataagaatgt actattgggc ttataacatt 28140
ttaaatgtaa tagtatagtt acatcacaaa aggggagaaa agggaataga gctatataag 28200
agtaacattt ctatatctca ctggaataaa attggtgtaa atctgaagct gattctgata 28260
agatgtatct gagaaggcct agagcaacca ctaagaaaat gattttttaa agtagtaaaa 28320
aaaaaatcct taaagaaatt taaatgctat agtagaaaat attctcttaa tgcacaagaa 28380
agcaataaag gtggaataga ggagaagaca acatgggaca tataggaaac gaaaagtaga 28440
atggtagaca taaatttact gtatcaataa tgacattaaa tacttttaat taaacagtcc 28500
aatcaaaagg cagagagtta aacaatcttt agttaaaaac aaaaagcaag attgaactat 28560
atactgtcta cagagccaca ctttatattc aaagatacaa atagactgaa agtaaaagga 28620
aaggaaaaga tataccatgt aaacagcacc cacaagaaag ctggagaggc tatgttaata 28680
tcagacaaaa tagacgttaa aaaaaaaaag tcaccaaagg taaagaggga cattttatag 28740
tgataaaaaa gacaatttat taggatgata taacaattat agacatatat ccagtaaaca 28800
acagagtgcc aaaatatgtg aagtaaaaac tgacaaaaat gaaggaagaa acagataact 28860
caacaatggt agttgaagac ttcaataccc aattttcaag agggatataa caactaggca 28920
gaaaacaata agaaaataga aggcatgaac,aacactaaaa gccaacaata cctagtagac 28980
attctcagaa cactatgaat aattaatacc aattctttac aaattcttca aagaatagaa 29040
gatgaaggaa cactttctaa ctcattattt gaggccagta ttaccctaat accaaaaatg 29100
11
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
gacaaaaaca tcacaaggaa ggaaaactac aaaccaatat tttttattat gcatggaaaa 29160
aatttctaac aaaatactag caagctgaat ccagaaacgt atgaaaagga ttatacacca 29220
gggtgggatt tatctcagga atgccctgtt ggtttaacat cccaagatca attaatgtaa 29280
tgtaccgtat caatagaata aaaaacagaa ccatgatcat ctcaatacac agaaaaatca 29340
tttgataaat tcaacacctt ttcatggtaa aaacattcaa aaaactagta ataggaaaca 29400
tcctcagtct gataagacat ctacaagaaa ctcacaggta acatcatact tgattggaaa 29460
agagtggata ctttttctct aagatcagga acaagacaaa gatgcccact cttgctattt 29520
ctagtcaaca ttgtactgga gtgtctagct ggggcaatta agtgagaaaa ataaataaaa 29580
gacatcccga ttggcaagaa gaaacaaaag tatctctatt tgcaagtaac ataatcgtgt 29640
atatggaaaa tcccagggac tccactaaaa aaccattaaa actaataaac aagtccagca 29700
agtacattag attcctgggg ctgccataac aaagtaccaa aaactgggta acttaagaca 29760
acagaaattt attttcttac agttacaggg gctagaagtc caaactcaag gtgttgggag 29820
tccctttgag acactgagta gaatccttcc tttccttttc tagtgtctgg tatttgttgg 29880
cttttcctgg tatttcttgg tttgtagatg catcactcca gggtctgcct ccatcatcat 29940
atgactgtct tcatccttcg tgtctctctt ctcttcttgt aattgcacca atcatattag 30000
attaagggcc taccctactc tagtatgacc tcaccttaac taattacatc tgtaatgacc 30060
ctacttctaa ggtcacattc tgaggtagtg ggagttagta ctccaacata tctttttgat 30120
gggggataca atccaaacat aacagcaaag ctacaagata caaaattaat tactaaaatt 30180
aattatgtat ctatatacta gcgttgaaca atctgaaaat gatattaagg aaataattcc 30240
atttataata gtattaaaaa ataaaatact taggaataat tttaataaat aaatacttca 30300
aaaacttaaa aaaaattgtt gaaagaaatt ttaagactga aataaatgga aggacattca 30360
tgatcctaga ttggaagaca aggttaaggt ggaaatcctt cgcaaatgaa tctacagatt 30420
taatgcaatc aatatcaaaa tcccagctag attctttaga gaaatttata ggttattata 30480
gaatttcaaa actaatatgg gatttcaaaa catacggaat ttaaagggac ccaaaatagc 30540
caaaacaatc tcaaaaaaga aggacaaact aggaggcact taacacttct caattccaaa 30600
acttactacc aagcaacagt aataaagact gtgtggtact ggcacaaagt tagacataca 30660
gctctatgaa acggaattga gaatccagaa ataaacctct atagttaatt gattttcaac 30720
aggaatgcca agaccatcta atgtaggaaa aatattattc gcaaaggatt cttaaatatg 30780
acacccaaat gcaagtgagt aaagtataaa taaattgggc tttaataaaa ctgaaacctt 30840
gtgtgcttca aagcacactg tctagaaagt gacaaaacag cccacagaat gggagaaaat 30900
attttcaaat tatatatctg ataagagaat tgtatccaga atatgtaaag agctcttaca 30960
atttaataag acaacccact taaaatggca aaggacttga atagacattt ctccaagaaa 31020
tattttcaaa tgaccaataa gcacatgaga agatgcttgg catcattaat tatcaagaaa 31080
tgcaaattga aaccagagaa gatacccctt caaggcccct aggatggctt gaatcagaaa 31140
gtcagataat aagaagtgtt ggcaaggatt tggagaaatt gaaaccctca cacactgctg 31200
gtgggaatgt aaaatggtgt gaccactttg gaaaaaactc tagcagtttc tgacaggtaa 31260
tcacagagtt attatatgac ccagcaattc cattttttag gtacataccc aagaataatg 31320
aaaacaaaag tcatggaaaa acaaaaacat ttctaataac actgttcata gtagctaaaa 31380
ggtgaaagca acccaaatga ccataaatta atgaatagac aaattttgaa tatgtacaca 31440
gtgggatatt acttggccat aaaaaggaat gaagtacaga agcatgctac aacttggatg 31500
agccttgaaa atgttatgct aagggaagga aaccagtcac aaaagacccc atttcattta 31560
tatgaaatgt ggtaaagcta ttgagacaga aagtaaagct attgagacag aaagtagatt 31620
atctgttgct tagggatgcg gtggatgaga ggatgggatg gcggtggggt ggcagctaca 31680
aggtacaggg tttctgtttt cttctctctc tgtcacgcag gctggagtgc agtgtagctg 31740
agacaacagg cacatgacac catgtccggc taattttctt ttttattttt gtagagatgg 31800
ggtttcatca tgtttcccag gctggtcttg aattcctggg ctctagcgac ccatctgcct 31860
cagcctccca aaatggtagg attacaggta caagccacca cactcagcct aggtacagga 31920
cttattttga gctgatgaca gttttccaaa atggattgtt ttagcagttg catatatctg 31980
taaatatact aaaaactcct gaattttacc ctttaaatgg atgaattagt acatgaatta 32040
tatctcaata aagctgtttt ttaaaaagag agtagacatg taaggttgct tggaaaattg 32100
actgcttgtg ttgtaaagtt aggccaagta tatctgtata cattccttgt gaacagtgcc 32160
attttagtga ttatataaaa tccaggacag aaacaataaa gtggtgggac cttatcacct 32220
agttctccag tttgaaatta aacctgttcc tcagttctat taactgtagc ctacattttc 32280
atgtcactga tcattatact tgggctattt atagtcttta gactgatgct cttggataag 32340
tttggcttag aagaaaaaaa aaagctattt tcagagtaat aagctctgtg agccagtagg 32400
ttaataagat tatagcaaca ccaaaggtat atttttaaaa atctcaatga catggtgatg 32460
ggaggggatc ttgtcagata atgatattct tgaggttgaa gcaatgttaa ctctttagcc 32520
ctagtacact cttttttccc tgaaatgttc aatgaatgct tccaaagaaa tggcagaatg 32580
ttttgttatt gttctgatct ttatgttgcg atgttacaaa tattgtaact tattttaggg 32640
actttgtaat tgtcacggca cccacagtac actgaatgct gtatggagca tgagcatata 32700
IZ
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
gaagacaggt acttattcat ctgggctttg ggctgagaat cgggagtcca ggtttctcaa 32760
gagttgggtt accgattagc ttagtccaga ctcatttgtt gagtaggagg attgtaccag 32820
atgcttcctg acagtcttat cacctccaac ttctgtgatg ctgaactcct gtgaaagatg 32880
gattatctct gtattttagg cagaaggaag ttagtacaag tactacatac atgtcacaca 32940
aaatcctttc agataattgg gtagcagtga catttctgga gattaactgg cttacacgaa 33000
ttgggccctg gaatgtttca tgggacatga aaactgactt gcttgatcct gtctttttac 33060
tacagatatg ctcctccacc tcttttacca gtcggtagtc ctgagagcag ggttttccaa 33120
actggatttt gtaaggtaat aaagtttgcc tgatgtcttt agtgcaggac ttctcagtta 33180
ttaacatgct aacttgagac aaccaataca acatttccaa aatgaagaga ctctgatgaa 33240
gccctttata tcatggagca catattaata cttcactgaa ccagaattca gaggaatcca 33300
gtttagaggc ctgtctgcct tggacacctc caaataggta taaaaattca ctgccaccag 33360
aaaacctggg cctgaccctg cccagtcccg ccttggtgct ccattagccc cacagttcca 33420
gcctcactga caggtcccct tctcctcttt gaggtcattt tctttttctc acacagattg 33480
aacattttaa gaattttaat aatgaaaacc gaattagacc tgttctgtgg ttgcactgtg 33540
ttgattcatc cttgcgttac cagtgcctgg cacagttcct gacattataa agcctggcac 33600
acagttatta ctcaagaaat gtttgtggaa taataattga atgaacaaat gaatgtgtac 33660
atgggggaca aaagccagct aatctctctt ctaatcaata tttacgcatt aagcttgcat 33720
agtataaatc agctctgctt gttaaggatt tttcccatga accagagctt caggtttgag 33780
atctgaaggg tgctgtcaca ccagctagtt tataaccacc tgtactggtg ccatgtcttt 33840
tattggtggc attggcctga ctctgcccca gcaccacctt ggtgctccat tagccctgca 33900
gttccagcct cactgacaga cccccttctc ctctttgagg tctttttctt tttctcacac 33960
agattaaaca ttttaagaat tttcgtaatg aaaaccaaat aagacctatg atgccccatc 34020
attataaatg ttctactcca agaccaacat cttctttgca cccttgaacc tgatactttc 34080
ccccatgtcc tcagaaggag aagcttttat gtattctttt tgagtttcca taaactcagg 34140
ctcagggcaa atggaaggaa cctctgcaaa tgcaaagata aatgtctcct gagctcttcg 34200
gttagggaca gtagggcaga cactgatgtt ggagcttcct ggctggctat atgccttcta 34260
caatcctcag agtccaatag gtatgctgta aatactttca ccctactgtc ttctgaaatt 34320
tgggaaaata ggtattcacc catctatgct cttctttcaa caaagatggg gattatttat 34380
gccttaagtc agctgtcaag tttaaacaaa gaacttcaac tacctccttt cttaaacttc 34440
ctaaagcact caagtcatat gaaaacaatt ctttaagatg ccatatttct tttaaataat 34500
cttatttaga agtattaaag gttattttga tacccctggc tcctttgata cctttcagaa 34560
gcttataaat tgttaaagaa ttacctagtt ccttttccag aaagtctcaa gatcattttc 34620
catgcaacag ccacatacgt tagggaccat tatttgtgtc agaaagaaat ttacgagctc 34680
cacaatagga tgtcctatgt agtagatcta tgtggtggaa aacaattgcc aaaggaaaag 34740
tcagctagat ttgagtgatg tggcacattt aacttatgtt gtctccattt tatatgccca 34800
cagtttgagt tctgccagct gaagttaaat catgtgacta agctggtttt tggtcttttg 34860
gttttcttaa taccaaggct ttttcaactt aacactgaat tccatataat gctttcatta 34920
agaatgccat gcattagact ctgcttgtca nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 34980
nnnnnnnnnn nnnnnnnnnn tgggacactg gttccaggac ctctacagat atcaaaatcc 35040
acagatgctc aagtccctta tataaaatag tgcagcattt gcaaataacc cacccacatc 35100
ctcctgtata ctttaaatca tctctagatt acctataata tctaataaaa tgtatatgct 35160
atgtaaatag ttgttatact atactgtttg gggaatcatg acaagaaaaa aaagtctgta 35220
catattcagt acaaatgcaa tcatcctttt ttaaaaaaat atttttgatc tgcagttggt 35280
tgagtccaca gaggtggaac ccacaggtat gcaagtcttg actgtgtgtg tgtgtctgtg 35340
tgtttatgtg tgtgtgattt ttttaacagt ttaagactca atagaagttg tagcaatagt 35400
acaacattcc tgtttaccct tcacccagct tcccccaatg ataatatctt acataacctt 35460
agtacattgt caaaaccagg aaattgacat tggtacaata ctattaactc aaatacagac 35520
tttatttgga tttcaccatt tttttttaca tccactgttg tttctgtttg tgtgtgtgga 35580
ggaatggtat agttctagga agttttatca catatgtaga tttatataac ctcaccagta 35640
caaccagggt ccagattgtt ccatcaccac aaagaaactc cttcatgtta ctccttaaca 35700
gccataccct gtcctcaacc ctaacccctg gcaacaatta atctgttctc tatctctata 35760
attttttcat tttgagaatg ttatagacat ggaatcatgt tgtctgaaac cttttgagat 35820
tggctttttt tttctttttt acttagcata gtgcccttga cacccatcgg gtggtatgtg 35880
tattaatctt ttgttcctat ttattggtga ctagtatgag caatatacat tgagcttgga 35940
tttgttggtg tatttgttgt taggttaaga ccaaagtcag aaagatcaaa tcagtagctg 36000
agtagattca ggaactggaa tcaaacaagg caaggttggg gctgggagag aggcagcagt 36060
acagagcgga gtggaaagca ggaactgaat tgagattgag agtgagagag gtgaggccat 36120
gctccaaccc agaccaggag agtgagaact gcagcagagg catgcccaga atcaactcca 36180
gtcaagaatg aggggttccc agaaggaaag ttgagcaggg gaagttggag aataatagct 36240
gagtgttccc tccacctgtc cctggcttac ccaggcctta ggggacactg ttctcctgga 36300
13
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
atttcctgat attgtttctg ctctgtagta tcttatgtta atattatgcc cagaatagaa 36360
ataaagatac cccgtggtca aaagttttat gcattttttc aaagtaaagc aaattcctat 36420
gcattgacct aacacataat gacacagtag cttccacaac cctgcagtgt ggaatgctgt 36480
gatctggctc aaattccaaa gctttttcat ggtggggaag aaggagagag aagaagaaac 36540
tgnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 36600
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 36660
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 36720
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 36780
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 36840
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 36900
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 36960
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 37020
nnnnnnnnnn nnnnnnnnnn nnnnnnnnna cccaggaacc cgaatccaaa tctttatcag 37080
ctgagttata ctgaagcttt ttgtttttaa tcccaaccta gaagaggcac tttttaccct 37140
tttttagtcc taaaatgata caggaaagtc aggagctcag tatgtggggc agagaaacag 37200
cttctaatct agagacaaaa ttcatggaat ccacattcat ggatatagct tctaattgct 37260
ttcttcttcc ttgcaatgaa atccagtcgt ttgtaaatcc tggttaggac atcttggtag 37320
aatggtcctt gagtttcctg tataaatgca caagtctcca atcaccctca ggcttctcga 37380
ccctaaacac aagctactga cctcttctag atagagtcaa ggtccatggc ccagaggtac 37440
ctactcctgc ccaattgtat ctcctcccca gtcctgcatg gggagggctg aactgttttt 37500
cactggatgg ttccttctgc cagtgcagcc ttcactggcc ttccccgacc tctgggactg 37560
aaccctgtgt ccctgtctaa tgctcgctac tttagctgac tcaactccat gcttgcccaa 37620
ctgcagtagc tgcccccacc cctgcccacc aggatgactc accctggacc cctccttacc 37680
ttggatccag tcagataata atatgtggaa ctcctttacc acctttaatc aggttggcca 37740
aaaacagtag cagaaaatgt gctcataaaa ttctgaatct ccttttggag atgatgtcgt 37800
gaaggaggta tggtgatctt tgaccaaatg ccccaagatg gtcggctccg ggccgcgctg 37860
catttcttat gccgcattgc tctctctccg ttcattttgc tttcacgata gcctggagtg 37920
aatatatctt cccttccttg tcagccctta cttctctctt aaattatgtt ctactgtgtt 37980
ctcacaagat tttctcgtaa aatataaaac caccttcctt ttgggagaat atgtagactg 38040
tgggggtttc agtagttctg acatcatgtt ccagggnnnn nnnnnnnnnn nnnnnnnnnn 38100
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38160
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38220
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38280
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38340
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38400
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38460
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38520
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38580
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38640
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38700
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38760
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38820
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38880
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38940
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39000
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39060
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39120
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39180
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39240
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39300
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39360
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39420
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39480
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39540
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39600
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39660
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39720
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39780
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39840
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39900
14
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 39960
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 40020
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 40080
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 40140
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 40200
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 40260
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 40320
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnt actgatgtga cgagatgggc 40380
tcagatctgt atgactaggg cactgtgctt agcatctaga catgatgtaa tattcacagg 40440
aaccctctta gttgtagggg gcagggtggt gaaaaactgg gaattaaggg tattacaaag 40500
gcataagtag gtatatttta tctcagactt gtcagttagg attttgtttc taaagccgac 40560
agaactactg gatcaaagaa gatagcataa aactgttatg tttctcagta gcactacggg 40620
aatccatcaa ctgcatcaaa ccttccccaa ttaacaaaag cttgggtaaa tcctgcatca 40680
actgaagact ggcattcata atgcccttaa ctggttccat cacattaaac ccattaagga 40740
agctcacaga cactgttgat gctgcttgga aaccagtcct agcaacaata gcacgtggaa 40800
tttgaggggc ttgagaaagg cagccctcga ggaattagct tgctaggtag ccagatcatt 40860
tggccagctg tggaaaccat gttagtggaa tagtggtgtc aatttgtgga gaaagattgt 40920
gtctggtagt aaaaaagtgg tttccccaaa gaaggtcaga gatgtcttct agagggctgg 40980
gtggcagcta ttagaagtca aactatacaa gaaagatatt ggtaaaaaat aataaataaa 41040
taaataaata annnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41100
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41160
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41220
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41280
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41340
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41400
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41460
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41520
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41580
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41640
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41700
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41760
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41820
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41880
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 41940
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 42000
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 42060
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnta gctgtgttaa 42120
tcaaagcgct tattttgtac aataataatg accttgaatt tcttaaaaaa aattacaata 42180
agctacaagt atcaaagaag cgaagttatc tggagtagtc tatataggag ctctttggac 42240
tttcttttat tatgctaaaa tagtggtgct tttaggattt acattattgt actctccaat 42300
acaaagtatg gggcatgtta aagtatacag tacaccattt tcatacatgt acaacattgg 42360
tggatgaaga atgtctctta gcagtaatac tggatgtagc ctctggtttt accagctgca 42420
tactctagga ctattatata agtaaaaatc tctcttgtga tactggaaag tgattagaat 42480
gtgcaaactg atatagtagc tttcatccgc ctcttaaagg gtaccaccac aggaaagtcc 42540
atttaagatg ttggtaggtt taacaaagtt ggaatgctgg cactgttgaa ttgggcaaca 42600
gttcttcaga cetggctcag agctacaatg catttagtat attaaagcag ctgacatgat 42660
gactttttgc gagccttccc aggcactgga gtttttctgt taatttgccg cactaggtca 42720
taaaagatct cattaacatt tatttttgat tttgcagaag attctaagaa tgcacagttg 42780
ttccattgtc ttgctagatt ttgaccttgt tccttcccta caactctttc atcttccaag 42840
tcacacttat taccaacaag aatcattgga acatcatcag tgtctttaac tcgaagaatc 42900
tgttctctca ggtcttgtaa atcgttaaat gtggactgtg ctgtgatgga ataaactaat 42960
gcaaaccctt gtccattttt catgtataaa tccctcattg ctgtaaattg ctccgttcct 43020
gcagtatcca agatttcaag catacactgt tgtgcatcta cttcaacttg ctctctataa 43080
gaatcttcta tcgtaggatc gtatttttct acaaaaattc cttgaacaaa ttgtacagtc 43140
aaagcagact ttccaacgcc tcgtgagcca agaacgacta gcttatactc acgcatgatg 43200
caagcttgtc aaaacctagt actctgcgaa cctctcacgc tgtcaccggg tctctgcagc 43260
cagcgtcgcc ccgcgctccc cgcgggtcgc tactctaggc gccacggcgg tcctgccgct 43320
gccgcgccgc tccggctggt ttacacgcct gaatctgggc gaggttttct caatcctatt 43380
ttatgaaatc catctgcctc tggtgtttat gttggggtgg gagttaataa tgaaagagca 43440
aaataatctc ataatgtact tgttcccatc aggcctgcaa tgtgtgtctc tgatggaaca 43500
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
agaaagagag agatcctgaa tcttctttgc catggcactg ccactcccac ctcgggccct 43560
aagcacagac gatgctatag accaggaatc accaagcttg tctgtgaagg cccagatagt 43620
gaatacgtca aattttttag gccacctgtg gtctctatca cattttcttc tttactgtag 43680
ccattttaaa aaatgtaaaa aagaatgctt agcttgtacg ctgcacgaaa gcatagttcg 43740
tggacccctg ctacaaacct gcacattctt tgttactagc taagaatatg tctccaaatg 43800
tggacatggt ctcctatcct gataagcaag ggtcctcctg ttcatctagc ccaacgctga 43860
tgctggaggc aggctttggc tcctgcttca tgccctcttg gccagcacaa gtgctgatac 43920
cctgtttact gctgttgctg tgaatgttaa agtctcagag ccaagaatct catgtgttct 43980
gctggcaccc gtgagcctgg cgggctaact gactaaattt caggtaaggt gaaatctcag 44040
acccctcaca gttcttgaga gaggcagaag gggaagggaa ggagaggggg catggtgaga 44100
gataccagaa aaaacactac acggaaaaga gatgacccaa atgaagaggt cagttcccca 44160
acccctacgc tggcctggtg tttaagtcca gtttccatgt tggccttcac agaatcagag 44220
ctcttctttg ctagataaag tgggggggga gtctttggct ccccctggag gctagagttt 44280
gccaacgaca ggtaggacct gaaacacaat atgaattgct ctttactgta aaacaaaagg 44340
aaagctgacc catgacgaga ttttactcaa atggcccagg ggttaggtat aattcaatta 44400
aaaagggaat ttgtccttta aagtaggggg atatatatat atggctttaa ctaaaaagaa 44460
aaatggacac agaatctaac ggtgaatgtt taggtattag tgacagtcat cattgttaaa 44520
gtgcagagag aaaattagat ctaacacaga agagtaacat ggtgctaaaa tttcccgcat 44580
atcccagaca acttcctgtt tgcagctcga tgtgctgatg agatactttt aacacgaatc 44640
ctgagggcag tggaccttag aaaaattatc atctacaatt gtgtgttaaa ttgctaagta 44700
attaagatgt aagatctaga aaatcacata caatttattt ccaacagctg aaattagaac 44760
tttctaaata aaagctaacc atacaaccca agggtttaac aagtcagagt ctgtttggga 44820
aagccagcgt gtgtgtcccc gagaggtgga ttgcaatcag taataaaggc atgagtccag 44880
ccctgtcaga tctgtctgac agcccctact gcactctgtc agaatctccg catggggatg 44940
ggtcaggttg cagatggtga cagcatcatc cgaaagtcta tgagaaagtg aaggtaacac 45000
aaagccctgg gatccctacc ttccacactg ctctgcttat ctgcacacac agctgtgagg 45060
ctccagccag gcccccagga gcaatttgtt atttgtgaca accttgtgtg catggtgttc 45120
tccctcaccc cccaccccaa cacacacaca gggagccaaa gtttactacg tttagtctaa 45180
gttgggtttt gagatttaaa tgttcaagac ttaagtgatt accgatgggg aaaataggaa 45240
gtggcctgga ggttctcagt tcgggggaga atggttgtgc aaatagaaaa ccatggtagt 45300
gaatctgcta cccagttttc taagaagagc tgcggttttg tttcatcctg tctcagagct 45360
ttattaatca gccatattgt aggtgacatg ctccagaaca cacagaattt atgtccttgt 45420
gctctttcct ctaagagcta attaatgtat ttttattggt gagaaagttc actccacata 45480
tttcttctcc ttgcatcatg atcaattctc caatcttaaa accaaatcac ccaccaccac 45540
cacccataaa caccctctgc ttcaatcctg gtccccattt acttaattcc catctttccc 45600
ctacttttaa aatctattca ttaaaacaaa acaaaagcca aaaacatctt tcctgggatt 45660
gcctggagaa gacttgacag gaccaagagg gaaaactttc taacccccaa tcactgttta 45720
gtttatggct ggaattatgg gggaaaagta gaatgaggtt gcttttgatg ttcttttgaa 45780
atgatttgca gcattgaaag aaatattagc aattaagcca ttttgatttt ctttaaaact 45840
gaaagtagag gccggcaagg tggctcacgc ctgtaatccc agcactttgg gagaacgagg 45900
tggatggatc acaaggtcag gagatcgaga ccatcctggc taacatgatg aaaccccgtc 45960
tctactaaaa atacaaaaaa ttagccgagc acggtggcgg gcgtctgtag tcccagctac 46020
tcgggaggct gaggaaggag aatggcgtga acccgggagg cagagcttgc agtgagccga 46080
gatcacgcca ctgcactcca gcctgagcga cagagcgaga ctctgtctca aaaaaaaaac 46140
aacaaaaaac ggaaagtaca gaagtgtata tatttaataa tcaattatat taatcaatat 46200
attatatatt aatatataca aatatattaa tctatatata tatgattctt aatgctgtat 46260
ttcttatagt ctgggtatcc cagtgaatga aatattttca gcagtgtttt gggaagtaca 46320
gagatcttcc cttatatgtt ccaggacccg ctagtgatac ctagaatcgc agatagtacc 46380
gtcccctata tatactgttt tttcaatata tacataccca tggtaaagtt caacttacaa 46440
attcagcata gtaggatatt aataacaata actgataata aaataataaa ctagagtaat 46500
tataacaata tactataata aaagttacct aaatatggta tttctgtctc tcaaaataac 46560
ttaatatttt cagaccatgg ttggctgcga ataactgaaa ccactgaagg caaaaccgca 46620
gataaggggg aactactgta tgctattcca gatatgtgtt attatgtccc aaactatcca 46680
aaatcttagt ggctttaaac aatagccatt ttattgtatc tcacactcct gtgtgaattt 46740
ggaacttgga caaggcttgg ctaggtgatt cttctgttct tggtggttta aacagaggtc 46800
actctggtat tttgctggag ggtctaagat gactgacatc tgggcaggaa tggctgaaag 46860
attgttttct tgttctgttc ctctaaaata ccagttcctc ttagatattt atgctttgtt 46920
tgttcactgt gctgatatag aggagatttc tctgaaaaga aaaCCatact ttcattgttt 46980
ataattatgc ttaaacttaa gaagataaat acatttttca taagtctgtt tctcatgcat 47040
tttgggccca gtggggacag taggtgagag cacctatctg tgacctcttc agcatatcag 47100
16
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
tctcagggta gtcagatgta cagatggggc ctcagggctc caaaagcaag tgctatggta 47160
gacaaggtgg aaactgcatg gccttttatt acctaatctt gaaagtcaca tagcatcact 47220
tctgtcatac actattcatt caagcagtta cccagaatca aggagaggca acatagactt 47280
caccttttca tgggaagagc attgcagaat ttgcagctgt gttgtacagt caccaaacat 47340
gcacacagga gacatctcca ttctattaca tgttagagta aatttctttt ctttttcagg 47400
tgtagtattt ccaaaatatt gttcccacca ttgaatttta tagcagtggc tcccagtctt 47460
ttggattttt aattccaatt ttgaaaaaaa agaatgagca agattgacag agggaagccc 47520
acctttattt tactaaggaa gaacatttaa aaataatgat tgcaataaca tgcacactac 47580
tctttatact cactatcatt ttataaaaca aaaggcatgt tcacatcaaa ataataggaa 47640
gggcatgttc ttaaaggacc ttttttaaat tgaataaatt tatctttatg aaaaagacat 47700
atcattttcc ttgtttttct tgtttaactg caggctggtg gaaactgtct catggagcct 47760
gcctttttgg cagattggat ttgggaatta ttcttctaga caaaaggtct caagtatgcc 47820
acttatactt tcttgggaca ctcaagattt tatggtcata atttcatttt tgagttagtt 47880
ctgtcaattg catagcttta tgtgaggagg actgcagttt gagttcaact ttctaaatta 47940
aatcaacgag gttttattga ttttcaaaat gtgcctacaa tagtgtagcc tcatggagtt 48000
tcagaaaagg atgacataat acatagttcc tgccaggagt caaattgaaa attaattata 48060
gagataggac tgatgcttat gaaatgctga taaataatgc aagttagaat gtgggtgaga 48120
atgtatagga gttacccacg caaactatag tagtctgggt gtcttcacta tgaatgtgga 48180
gttgaagttc aaggtacacg atttggatga aaggcaaaag gagcatgttg catatatgat 48240
taattgaatg aggattcgtg gataagaatg aatgaacgaa tgcagtgtgc ttaggggcca 48300
tttagggatt ctaggataca atggttttta acgtgcctta tcttaggttt cagagcagaa 48360
aacaaaatgg attcagcatt gttttttgct tgtttttgag tcacaatata ataattgatt 48420
aaaattccta ctttcttctt actctcaaag tgaccatgtt tttctctgag aagcggatgg 48480
ctgactgctc attctagaca taaatcatta atagtcataa taaaaagaat cagctactaa 48540
gtaacagctt cagtactgta ggtcagcacc tgaatatctg ggtgaccttt cctaaaatgc 48600
cgaagagtgc ttctgacagt ttgggatcga ggagccctcg tgtgatgctt catcactaat 48660
gacaaaataa atctcaagtg tcaagaagtg ctgccctgaa ttcacctcga ctttgccatc 48720
agcaatggat taggggcaac atccattagt gcaaggtaga tcccgctatt tattcactca 48780
ctgctctggg ctgacaactt aggtgtctgc ctgtttcagg aaacaaaagg tcacatagga 48840
gtatcagtaa tggcctgctg gattttctct ttagacattt actggaatct attatgatct 48900
agtctggttg ttttgatatc tgccatctga actttaaaaa caacaacaga aaaaaaaact 48960
tacagataat tttttaagag agggatcttg gaggctttgt gcaattattt tcctatgaat 49020
gttgatcaag cactgtgtga tctttgagtt cattttcagt cctgttttcc tgccatacac 49080
aatcatgttc ccatcatcta gaaatgcacg gacttaagca gtgagagtcc ctgcccctcc 49140
aggcctttct gttgctgtac aatatcaaaa gatttaatat gaacatagat caaaaattgt 49200
tttcttgttc tgttcctcta aaataccagt tccttttaga tttttatgct ttgtttattc 49260
actgtgctga tattaatata aagaagactt ttctggaaaa atatttttta aggtagttat 49320
gcttaaactt aagaagatta atacatttct cataagtctg tttttcacgc attttctgac 49380
aagacacata gtagcacttt tcttccaaaa tagcccactc ttccaatagc ataagccctt 49440
tgctatagtc tattacatct gcggctgtgg tttcaaggca atcatgtcca tattaatata 49500
cttgatgctt ttcttctgtc ttcactgatg ccctgtgctg atgcattnnn nnnnnnnnnn 49560
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 49620
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 49680
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 49740
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 49800
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 49860
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 49920
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 49980
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50040
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50100
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50160
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50220
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50280
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50340
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50400
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50460
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50520
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50580
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50640
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50700
17
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50760
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50820
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50880
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50940
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51000
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51060
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51120
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51180
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51240
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51300
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51360
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51420
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51480
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51540
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51600
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51660
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51720
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51780
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51840
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51900
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 51960
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52020
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52080
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52140
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52200
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52260
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52320
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52380
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52440
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52500
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52560
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52620
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52680
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52740
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52800
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52860
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52920
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 52980
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn ctcccttgga 53040
actttcctcc gagatgcttt tacttggagg cttctcattc attattcaaa acccagtaca 53100
accatcaccc tgtatgtgaa gcttttcaga attccctaga actaattaag tagtctttcc 53160
ttttttcccc ttaagcataa tgtactcacc ttaattatag cacatgtttc attctgtggt 53220
aattactttt tgcatgtttt taatctccca ggaaaccaaa ggttcaaagc cccggcttca 53280
tattagaatt tcttggaaag cttttaagaa atactgtcat ctgaacccta tgccaaaacc 53340
ttggtgttaa gcttcacatt gcctcttgga tagagaagaa atagcaaatt atagataaag 53400
aacctctggg tgtggcctgg catcagtgtt ttgtttttct taaagctctt cagatgattc 53460
cagtgagcag ccagagtctg aaaccttgct gcactaggct gtgatctcct ggagagtgaa 53520
aactgtgtcc tattcaagtt tgtggacctc tcctccctcc ctaccctcat actttgcaag 53580
atgcctggtt tatagcagga gctcaagcaa gagctgtgga attaatgatc acctgcttca 53640
tttgttcggt cagtgattgg gattattgaa gattaggttc agtctattac cacttgtatt 53700
agtccgttct catactgcta taaagaaata cgtgagactg aataatttat aaagaaaaga 53760
ggtttaattg actcacagtt ctgcatggct ggggaggtct caagaaactt acaatcatgg 53820
cgaaaggcac ctcttcatag ggcggcagga gagagaatga gtgctgagcg aaggggggaa 53880
gccccttgta aaactctcag atctcatgag aactcactga ctatcatgag aacagcatga 53940
aggaagccac tcccatgatt caattatctc cgcctggtcc cacccttgac acatggggat 54000
tgtttcaatt caaggtgaga tctgtgggga cacagagcca aagcatatca ttcgacccca 54060
agcccctccc agatctcatg tcctcacatt tcaaaacaca atcatgccct tccaacagtc 54120
ccccaaagtc ttaattcatt tcagcattaa cccaaaggtc caagtccaaa gtctcatctg 54180
agacaaggca aatcccttct ccctatgagc ctgtaaaatc aaaagcaagt tagttacttc 54240
ctagatacaa tggggttaca ggcattgggt aaatacacag attcccaatg ggagaaattg 54300
~8
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
gccaaaatga aggggctaca ggccccatgt gagtccaaaa tccagtaggt cagtagttaa 54360
accttatcat tccaaaatta ttttctttga ctccatatct cacatccagg tcacactgat 54420
acaagaggtg ggctcccaca gccttgggca gctctgcccc tgtggctttg cagggtacag 54480
ttgctgcttt catgggctgg cattgagtgt ctgcagcttt tccaggcaca tggtgcaagc 54540
tgttggcgga tctaccattc tgggttctgg aggatggtgg ccctcttctc acagctccgt 54600
taggcagtgc cacagtgggg actctgtgtg ggggcttgta tcccacattt tccttcctta 54660
ctaccttagc agagattctc catgagggct ctgcccttgc agcaaacatc tgcctggaca 54720
tccaggagtt tccatacatc cttggaaatc taggcagaga ttccccaacc ttaattcttg 54780
acttctgtgc atccccaggc ccaacaccat gtgaaagcca ccaaggcttg gggcttgcac 54840
tctctgaagc aatagcctga gctataatct ggactctttt agccatagct ggagctgaag 54900
cagctgggat tcagggcacc atgttccaag gctgcatagc agccagtgaa acaatttttc 54960
cctcctaggc ctccaggcct gtgatgggag aggctgccat gaaggtctct cacatgtcct 55020
ggagacattt tccgcattgt cttggtgatt caatggagaa atctccattg aattcagctc 55080
ctgttactta tgcaaatttc tgcaccaggc ttgaatttct ccccagaaaa tgggtttttc 55140
ttttctatca catcatcagg ctgcaagttt ttcaaacttt tatgctctgc ttcctcttga 55200
acactttgtg gcttagaaac ttcttccacc agatacccta aatcatcttt ctcaagttca 55260
aaattccaca tatctctagg gaaggggcaa aatgccacca ttctctttgc atagcaagag 55320
tgacctttac cctagttccc aagaagttct tcattttcat ttgagaccac ctcagcctgg 55380
acttcgttgt ccatatcact accagcattt tggtcaaagc cattcaacaa gtctctaaga 55440
agtcccaaac cttcccacat cttcctgtct tcttctgagc cctccaaact gttccagcaa 55500
ctggctggta cccagttcca aagtcatttc cacattttcg ggtatcttta gagcagtgct 55560
tcactaccca gtaccaattt agtgtattag tccattctca cactgctatg aaaaaatacc 55620
caagactgag taatttataa ggaaaagagg ttggccaggc atggtggctc atgcctgtaa 55680
tcacagtact tgaggaggcc aaggcgggca gatcacctga gatcaggagt tcgatcccag 55740
cctggccaac atggtgaaac cccatctcta ctaaaaatac aaaaagtagc tgggcatggt 55800
ggcacacacc tgtgatccca gctatgcagg aggctgaggc aagagaatcg cttgaatcca 55860
ggaggcggtg gttgcagtga gccaaggtta caccactgca ctccaggctg ggcaacaaag 55920
tgagaccttg tctcaaacaa acaaacaaac aaacaaagac atttaattga ctcgcagttc 55980
cacagggatg gggaggcctc aggaaactta caatcatggt ggaaggcacc tcttcacagg 56040
acagcaggag agagaatgag cactgaccaa agcgggaagc cccttataaa atcatcagat 56100
ctcatgagaa ctcactcact atcatgagaa cagaatgagg gaaccacccc tatgattcaa 56160
ttatttccac ctggtactgc ccttgacaca tggggattat tacaattcga ggtaagattt 56220
gggtggggac acagagccag agcatatcac cacttttctc cctcagcaca gaacttggag 56280
taggagaacc tcgcaaggga tggggatccc attggtgtta aactgatatg caatgtccat 56340
ggtacaccac aaggtgattc gctttaatgt tcttgattcc taagactttt aaaaaattgc 56400
ttcaaataac aagggggcct gcagagccat acatgcacaa aacactgtaa tgtctcctcc 56460
cctgtctgat gggcatgctg tacaacagcc atttgaggaa cgacattaga gaatagattg 56520
ggcttcatta ttaggcctct tcttctacag tttgctcatt ggattcagga aaactgtctt 56580
aggcccgcat gtgcttttta gtgcccaggt gaacttaggt aacaagtagc actattgctt 56640
gacatggcat tatctgccca tgtttggtct ctgcttggac cagcaaactt aaatgtcaga 56700
cttccctaag gtaaaagtca aagatatgct atagactgca tctgatattt acagttcttt 56760
ctagctaaga agagctggct tttgtggtaa atattctctc atgcaaatta gaaaggtttt 56820
cccccctttg tacatatgcc tttgatgtta ttactttctt tgcttttatt tgattttgtg 56880
aaagtccaca gatgcttttg aacacataat ggcccctact tgtggctact aaaaatagta 56940
attgaaaaag tgagggatca gttttagaag cctattaatt atgtatttaa atagtttcaa 57000
attgcaaaaa aaaaaaaaat atgtcttagg gatttagaaa attactcttt cagtgctgtt 57060
tcaggtatct tcttttcaga acttgagtta aggaaggtga aaagttgtca ttccactttt 57120
tctaccattc cagatccatc ttctttaatt cctgcttttt cttttccatc actattttat 57180
ttctccgctt ttctctttac gtgatgccaa tagtaatatt aataattgtt ggtcgggtgc 57240
agtgactcat gcctgtaatc ccagcacttt gggagctaag ggcaggtgga ttgcttgagc 57300
tcaggagtta gagaccaggc ctgggcaata tggtgaaacc ccatctctac gaaaaataca 57360
aaaaattagc tggacatggt ggcacgcacc tgtagtccca actatttggg aggctgaggt 57420
agaaggatca cttgaacctg ggagacagag gttgcagtga gctgagatca tgcccctgca 57480
ctctaacctg ggtgacagag tgagaccccc tctcaaaaat aataataata attgttgctt 57540
attggacaac tgtgtgtttt acacagtact aagcacttta tctcatttaa tctctcaaaa 57600
ccctgtgaag taggtactgt tattagcccc actatacaga ggagaaaact gaggcagcac 57660
aaggaactta agtaactcta ggtcacacag ctaggaactg gttggggaag ggttcagatc 57720
taagccatct gttttaagag cccaggctct taactatttg ctacctgacc caaagttcag 57780
agtggtaaat ggtagtagaa tgaagactaa gcatactgca tttgatttaa aaaataataa 57840
agttattttt cccccatctt aacccccatt actttaataa tggattcact tctcagcact 57900
19
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
attaatgaaa gaatttaata gaacaagtac tgagcttgag atttcatgct atcatttcta 57960
atcagacaca gaatttgtaa ccagtaatga caaaaaatag caaattgtgg ataacaagga 58020
gatgttacag cctacttagc atcttgggtg gggggcaggg acagctggag agggacagtg 58080
gtccctggag tcagccttct atggactctg atggagtctg gccaagggaa gaccacactc 58140
agccaaccat tttggggttc caggagcagg gacaacaggg cagcagctgt gaaagcttac 58200
acctcagtga ttaggtgggg agggtccata agaggccaaa aagggagaga ggcaccccac 58260
tcccaccccc atgcacacag aaaggaaagg ctgggccaac tttgcaggac tgactgaccc 58320
aacatctagc ccaaaccaag catttgacac aggcggaagc tagaagactg gcctgagatc 58380
ttaagtatag ttagagggag actagagcta gtccagggta ccttggcacc tttgtccaag 58440
gatttttcta gcataaaatg aaaaaaaaaa atgactcttt gaggaaatag gagagacgga 58500
gcttacactg atgtgtaagc tttcacagct gctgccctgt tgtccctgct cctggaaccc 58560
caaaattgtt ggctgagtgt ggtcttccct tggcttccta gaattgacct taagaggata 58620
tcattttcag tgttgccttt ggtaataaaa caaaccaatt tgcagtgact attccagtag 58680
ctgatttctt taagcacaaa gcatactttt aggggtcttc agaggcatgg tagaaacccc 58740
cttacctgac taaatcaggt ctggtaatta gttggttagt agaaaagttg gtaaatgcag 58800
gaaaacatgc aaaaatataa atatgtatgt aaatatggtg ttttaaccta taagcataca 58860
ttcgtttgat catctttgga tgcagagtga gagatttttc ctttgagttg ggctcttcaa 58920
actggagttc cccattgttt tctttgcatc actacagtca ttctgcagct ttgattttta 58980
gtcttaagtt tctttaaagt ggagtgagac cataaaaaca catgtgcctc aatctgagca 59040
caggatcctt catcttaatg atttttctca gtcttttttt ttgtattgct cacattgcaa 59100
tgattttctt ttgcaagtca tcttccccaa gtctttctta cattcacttt catagaaata 59160
gctccctttc ctttcacact cagatttcat tatttatgtg attggtgtta aattaaatgc 59220
ataattaata acaaacagaa ctggcataaa aaagttgggt acaaacatag gtaactcttc 59280
agaaacataa agtgctgctg gtggaagcag aaggaagatg ggtgagtgaa agggaaacac 59340
ttgcagtgat ggtttttggt gtgtttgtgg gaggggtggt tactttattg ggttggtgaa 59400
atgaaaactg gccaactgat gttaaaggga aaattatttg taatgcttgt taaagcatag 59460
tcaggaccat cataacaggt ctagggacca ctgcaatggg ttttgcagta taaaagagag 59520
atcaggctca actcctagta caaaaaggaa aagtgtgaat ttagagccaa ggtccagggt 59580
gggagagtta gtggatagaa aattaccaag agggatctac cgccatatca ccctgaatgc 59640
acccagtctc atcagaaaat taccaagagg aaacattagg ggtaaagaag actttgggta 59700
agttgaccta actggattct tgctgaagac agagcagggt gatcagatat tgcccagggg 59760
atctgatcag atattgaaga caaccagcca tctagagtcg gggattcgct gacttggcag 59820
gattcttgct aaaattgggc aatgcagaaa tgaacacaga agtccatact tcatagtcta 59880
gttgagaaga gtggtccaaa gactagggtt tggtcaagga gagattgtgt gtcactgggc 59940
ctctattgta gacacacctc agatttgtgg aagaaactct acctttggga aaacaaatgc 60000
acctacttct tatgaaagaa gtaataacat ctaaaaaaat gaagcattat ttgagtaaca 60060
aagctttatt taatgtcagt ccatattatt cttttcaacc atttccagaa atgtccttat 60120
cattcaacaa aaatctactt ttatttttct ttgactcaaa aattttgttt taacctaaga 60180
agatctgcat ataaaaagtg attgtgattg tcacctcatg gacattatca attgtgtaag 60240
tcaattctca cactgctgta aagaaatacg tgagacggac taatttataa agaagagagg 60300
tttaattagc tcacagttct tcaggctgta caggaagcat ggttgggggg gcctcaggaa 60360
acttacaatc atggtggaag gcaaagggga agcaggcatg tcttacatgt ctggaacaga 60420
aggaagagag agaagaggga ggtgctacac acttttaaac aaccagatct tgaaaagcaa 60480
aagggaaatc tacccccatg gtctaatcac ctcccaggtc ccacctccaa cattgggaat 60540
tacaatttga catgagattg ggcagggaca cagacccaaa ccttatcatc agtgaattcg 60600
gaatactaat acagaatgtt tttcattgaa aatcagttta taaataaatg gtgcaaggaa 60660
ttctagagta atgcattagc catgcattac tgaagtgctt aagataattt ttaaaaactg 60720
atggactaaa agttggtatg agaaaaacac tatatatcat ttcctaaagt tgtnnnnnnn 60780
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnccaggtg tagtggtggg 60840
cgcctgtagt cccagctact cggggaggct gaggcaggaa gtaatggcgt gaactgggga 60900
ggcagagctt gcagtgagcc aaaatcgcgc cactgcactc tagcactcta gcctgggtga 60960
cagagcgaga ctctgtctca aaaaaaaaaa aagaaaaaag aaaaagaaaa cgagaaaaag 61020
aatgatacaa tggactttgg ggacttgtgg ggaagagtgg gaggggggca agggataaaa 61080
gactataaat atggtgcagt ttatactgct cgggtaatgg gtgcgccaaa atctcacgaa 61140
tcaccactaa agagcttact catgtaatca aaaactacct gtaccccaat aacttatgga 61200
aaaataaaaa ttaaaaaaaa gaagctgcca ggccacatct tgaatgcttt gctgcttaga 61260
aattacttcc accagatatc ctaaatcatc tctctcaagt tcaaagttcc acagatccct 61320
agagcagagg cacaatgcca ccagtctctt tgctaaagca tagcaagagt gacccttgct 61380
ccatttccca ataagttcct catttccatc tgagacctcc tcagcctgga attcactgtc 61440
cttattgaaa tcagaatttt ggtcacaacc attttaaaag cctctaggaa gcttcaaact 61500
2~
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
atccctcatc ttcccatctt cttctgagcc ctccaaactg tcgaatctct gcccattacc 61560
cagttccaac actgcttccc attttttggt gtccttatag caatgcccca cttcttggta 61620
ccaattttct gtattagtcc attctcgcac tgctataaag aaatgcctga gaatgggaaa 61680
tttataaaga aaagaggttt aattggctca tagttcttca ggctgtacag gaagcatggt 61740
tggggaggcc tcaggaaact tacaatcatg gtggaaggca aagaagaagc aagcatgtct 61800
tatatgtctg gaacacaagg aagagaaggg gaaggtgtta cacactttta aacaaccaga 61860
tcttgaaaag caaaagggaa atctaccccc atggtctaat cacctcccag gtcccacctc 61920
caacattggg aattacaatt tgacatgaga tttgggcagg gacacagacc caaaccttat 61980
catcagtgaa ttcggaatac taatacagaa tgtttttcat tgaaaatcag tttataaata 62040
aatggtgcaa ggaattctag agtaatgcat tagacatgca ttactgaagt gcttaagata 62100
atttttaaaa actgatggac taaaagttgg tatgagaaaa acactatata tcatttccta 62160
aagttgttct gttttgttta aatatggttt aaaatggtgt tgggggaaaa gacatgtccc 62220
cttagggatt gtcagaaaga atctaaagca gcttagcaaa gagacagaaa tgtaaatgtg 62280
atatttatca ggggaaatga gggtatgatt aaataaaatg gaaccagcct ccatgtcaga 62340
aaatacctta atgaggaagt catnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 62400
nnnnnnnnnn nnnggtctaa tcacctccca ggtcccacct ccaacattgg gaattacaat 62460
ttgacatgag atttgggcag ggacacagac ccaaacctta tcatcagtga attcggaata 62520
ctaatacaga atgtttttca ttgaaaatca gtttataaat aaatggtgca aggaattcta 62580
gagtaatgca ttagccatgc attactgaag tgcttaagat aatttttaaa aactgatgga 62640
ctaaaagttg gtatgagaaa aacactatat atcatttcct aaagttgttc tgttttgttt 62700
aaatatggtt taaaatggtg ttgggggaaa agacatgtcc ccttagggat tgtcagaaag 62760
aatctaaagc agcttagcaa agagacagaa atgtaaatgt gatatttatc aggggaaatg 62820
agggtatgat taaataaaat ggaaccagcc tccatgtcag aaaatacctt aatgaggaag 62880
tcatacttga ggcctttgtg tcatcttctg aaagactaag agcctggact ggccaggaat 62940
ggccttcacc agaactatgg taggaagagt aagttgagtt tatttctcag gtcaggttca 63000
acctcctgaa atcccagctg aattaaagtc ttcattaaat tcatgaaagg acacaagagt 63060
cacagcctgg cctcttttaa catcgctttc ctggaattga atggaaagtc agcttctagt 63120
tctgtggaag gcaggtccca ggggaggtga gcagcttcta attccacagc tcctgcttgc 63180
ccttttgaga gctccagctc atctagcagg tcttgcctgt ttcacatctt tggagttaaa 63240
atactctgta gccttcatct cctcccctca aacatgtatt taaaaaaata caaactttaa 63300
gttaaaaatg tattagaaat gttgtctgtg ttattggaaa gcaccctcag attctaaggt 63360
atacacagat tacttggggg tcttattcaa atgcagactt,tgagtcagtg ggtttggagt 63420
ggcgtctaag agtctacatt tccagtaagc tccccagggc aagggaggga gggactggtt 63480
tcactggttc atgggtcata ctttgagaag tgaggatcta aaagacctga tttcaagttc 63540
tgacttctga caactgtttt cttacttgta aaatgagagg ctttggatga agataccttt 63600
taagtctgcc cgtctgcagc atctcaaggc acagagagag agaaggctgg agattgtcta 63660
gagcttgatg tctgctctga gttttcaagg actccccgtt ataacactgt gaatatagaa 63720
aggaaaagtg aatgcaaaca gggaaggaaa tacagtttgt gtctgacaat cccagagtgg 63780
ttacaattat tcacctgatg cctttgccaa catttaaagg tttggcttta attagatcac 63840
acaatccaca aatgaacctt ctaattgtgc tgtgtttgtc tccaataagc tgttgggtag 63900
gattaattct ttccacagaa tgcccaatgg aacagtttag ttttcaatac aaactaagtg 63960
tagtcacaaa agattactac ctttagatct taaaaatatt tttaaacaga gtgcaagtaa 64020
agatacagtt tgatatgaca ttttttatac taactgtgtc agtcatggag gaatttaata 64080
ccattttgaa atctttctta tccttcggat tcccctctga tgttctcttg ttagtaaaat 64140
cttcccagag gccacgtgag aattagtagc caccttctgc ccctctgctc ctatttacac 64200
tgctgttcgg gtgcacacca catggttggg cttgagctca tttagttttg tatctgtttt 64260
tgtccttctc tgaattacga gctccttaag ccatcctgac agagtaggaa aaaaatttgg 64320
atgacttgga gcacaggtac atgagtttac actttgttct gttctgatcc tatgatccca 64380
ggaggccatt ctgccttcgt ttttgataag ccactttccc cagactgtaa atcccatatt 64440
ctcaggggtc tgaaagtcca aatgccttca gggaaaggtg gtaggaggaa tgcagaaagc 64500
agtggatggt gatagtctag ggggcagtag gaatcttggt taacctgcct tgccccaaga 64560
tctcagatgc attttcttta aacacaacag ggttgagctc aagcagaaat taacacccag 64620
ctaaagactt tgcttcacat cacctggttt ctggtttgca cataacaggg gctcccaagt 64680
tgagcttagc agaaagaaca gtggaaatat tcaaaataat taataacaag gatagccagg 64740
gaatcaaaat agcatgttta tagctctaag gtttacccta tagcatcagg ttttaaccct 64800
ttagttactg gatcaaggga aagtccagga cttcccagtg gagaggccag ggcagttgag 64860
cctggtgaaa tggaatatca gccctgagaa agacccccag gctttagtca gagaaacatc 64920
atttcacaac cctggctagc cacttactgt ctgttagatc aagttgaata atctgtctga 64980
gcctcagttt tctcatctgt aaaatgggtt aataatactc actttgcagg actgtggtga 65040
ggattaagtg aaccaacata tggaatgttc ctcaaagagt gcctggcaca aacagatgct 65100
21
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
cactcagtga aagtagtacc aggggctgtt gtttactatt atctttaaaa gataataagt 65160
ggttattgat tgaatgcaga cagcttgtct ttgtgttact catgctttct gtagtcagat 65220
aattcaggtg gaccacaaga gatcttggtt tactgttgcc atcaaggtgg cttaccccag 65280
ggctcagggg ctgggtgggg gcagttctga aagacataag gagaagggga taggcgaggc 65340
tttgttccct gggctggagc tatatgaaag actcccaagt ctttagatca cagcatccac 85400
aggccgctcc tgatctataa aagctcactt tctgatttgt aggcatttta aaagacacag 65460
atattcttat aaagatgctt agctagtgat tatcttctga gaggcatgag cggcaaactt 65520
gagcctctta tgctgtaaat aagagatctg ctacataggc atttccctgg gtcaatcctt 65580
ctgcttttgg aatccctcgt gctgcactta tacattctct aaattgtctc caggacacac 65640
actgggtctc atgacaggag gaatctccat tgagaaggat gatctgtcca tagtcagcac 65700
catagtacag agaaaaatat accctccaca cccaggcaaa ggttgaccac cctattctct 65760
gtagtaaccc agcctaaata aagacagctt ttattttaca attgctcaca acattcacca 65820
ttcactgtag ctgagtgtac tcagctatct gcaaaacagc atccctgcat tattgatgtg 65880
agggtcgtga gaccttgcca agtgtttttt ccatattgct tactcttcaa cttgaaaact 65940
gattactgtg tcctgaaggt tatattttta aaggaaattt tcctcatgca aaatgctgga 66000
atgtattttg ttccttgttc acttgctgtt acatgtgtca ttgactccca acctctgatg 66060
tgtttctttg ttttctgtgt gttgcaggca gcatagtgta cctcgggatg atggtggggg 66120
cgttcttctg gggaggactg gcagacaaag tgggaaggaa acagtctctt ctgatttgca 66180
tgtctgtcaa cggattcttt gccttccttt cttcatttgt ccaaggttat ggcttctttc 66240
tcttctgtcg cttactttct ggattcgggt aaggttttct ccttcatatt ttatagtaat 66300
gtgtttattc ttcataattc tccactctag agaaccttat tgggaggaaa ttatcccttc 66360
acacatctcc ttccatatcc tcatgtcagc cctggaaacc acaatgctgg ttcttagaaa 66420
tataattatc aacctttaaa tacaggacag aagggaacac acacctatgt tttcatagct 66480
gattattact tctagaagaa gagtattgtt ttaaatggaa tgctgtttct atggtagaac 66540
acattcacac atatatgctg cttctgaaaa tttaacataa gtaggaaata gctctcgtcc 66600
tacctgttat gaactaatta aaggtttgag ccattcaagc atgctcttaa attcaggatt 66660
agagtgtctc aaataaatgc acttttattg gtctatgtta acttggcata tttaacttgg 66720
tagaaagcct ccttgagacg taatttttag taagttcctg acttctgttg aattaacaga 66780
agttgtgcta ttacccaaat gcagtggtag agtgtagccg atgttcattt tctttaaaga 66840
gcttaaagca cttataacta tactaaatga atccataagc atagaaaaac ctaatagctg 66900
aataaggaat atatgagaaa ctttgcttca ataagaaaaa aacatagtaa aaatataaaa 66960
tttttataac gaatgggtac taggcttaat gcctgggtga caaaataatc tgtacaacaa 67020
acccccacga cacaagttta cctctataac aaatctgcac atgttcccct gaacttaaaa 67080
gttaaatata tctatatata aaatttttga ttcatgatta agaacttaga gacttttttt 67140
tcaagatgtc taacacaaaa tgaatctgta ggaataatat acaattttta aacataaatg 67200
aagaaaatga gggaaacggg gctgggttat aaactttaaa agctatcagt tgtggttgtt 67260
agtgatacat caccacaacc tacaaagcag aacacagttt tgtgaaaaga ggtcagagga 67320
ctatagcatg tctttcttaa ttatgtatag tagcatttcc cacagctcct atgctacatt 67380
ctgcacacaa tgagcactta gcaaatactt gactaagttg cacaaatttt aggagctttc 67440
cttagagtga agacagtgca tttcagtaaa ccaagatggc tggagagttt gggaaagtgt 67500
aaaatatttt gttcttgggg aaaaaaaaat gttcagaatc aatcatttgc tttgtagcaa 67560
cggagaaagg ggagaggagg ctttttctcc acaggggagc cccttggtgc tcagtttctg 67620
tgagactggc tacatacgct gctgacttga agatggattt tggattatgg actcaaaggg 67680
tttcagcaga cctaaatgtt gaatgattga atgaatgaat gaagaacaga tgttacttgg 67740
ggctattagt taaaggacct tctcagtgag taggtccata ggataagata ttcttgccac 67800
caggaaaggc caagcttcct tctccctttt attatctata ataaaaccac tagcctgtca 67860
acagaaggct aacagcataa gcactgaatt ttccagtcct ttcagtttgc aatctttttt 67920
gggaaattga agcaatatcc aaagacccaa aagtacatga ctgtggtcaa attattgtgg 67980
caattttaat tgccaatctg gattgagaca tggtcgtccc aacagactgt agagaaaaag 68040
gtccattgtg gtggctgcaa aatattccaa accaatggcc tgtcccagaa gcgaattcaa 68100
gggcaacata aaagcagaga caacttagat gtattcttga tatantcctc attcctactg 68160
cagacaaatc attcttcagc caacaggtca ggactcccgg gggctgggaa tcaagaaatc 68220
ccaggtgtgc cctactcctg cccattatca tggagtctta ggaaagtcat gcaggctctc 68280
ctgcctgctt tttcttttgt aaaagaagac attaggactt tgcattgtta ccttggagtg 68340
tgaagatcaa atgagatgat gtattgaaag tacaaggtag agaagataat gtaggataat 68400
gtttagaaac actcactcat agtttggcat ttcttatctc ctttgttcct cctcacatac 68460
attttgctaa gctgttctat gttgattgcc ttggagtgac agcaggggag gaggcagaaa 68520
gagactcctt tctggctttt taaatttcaa gtctggccca tgcaagaaac agaaaggcct 68580
aatcagggac aaaatatttg ggagaaaaca tggcaagtaa aatcaagcat aaatcaagtt 68640
aatctctata ataaatggtg agtacctagg ctggggacag ggctgagggg ggagtcatgt 68700
22
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
gagcttagtt accttagtcc aggtggcaaa atctctgcag agttttgtgg gatccagcgg 68760
ctgacaggac ctgagaaatg aagttattaa ggaaaatgag ggaagaaatc tatattggtc 68820
agcattctcc agagagagag aaccaatagg aaagaatgga tagaatagat agacaggcag 68880
atagatagat agatagatag atagatagat agatagatag atagatagat agatatgaga 68940
caggactaat taaggcaacg gctcacacaa ttatggaggc tgaggagtcc catggcaggc 69000
cttctgcaag ctggaaatgc tgggatgcca gtagcatggc tcagtataag tttgaaggcc 69060
tcagaaccag ggaagctgat aatataactc tcattccaag gctaaaggcc taataactca 69120
gggtgctgct agtgcaagtc ccaaagtcca aaggccagag aacctgggga tctgttgtcc 69180
aaggacagga gaggaaggat gtcccagctc cgagagactg caaattagaa tttcctctgc 69240
ctttttgttc tctccaggtt cacaaccgat tggatggtgc ccacccaccc ctgagggcag 69300
aacttctcca ctcagtttac caatttgcat gcccatctcc tccagaaaca ccctcacaga 69360
cgcacccaga aataatgctt tactagctgc tatagtttgg atttttgacc tttccaaatc 69420
ttacgttgaa atttgatccc cggtgttgga ggtggggcct gatgggaggt ttttaggtct 69480
tggaggtgga tctatcatga atagattaat gtgagtgagt tattgctcta ttagttctca 69540
tgagagctgg ttgtttaaaa ataggcctgg caccccccag ccctctcttt tgcttcctct 69600
ctcactgtgt gttctctgca catgccagct cccctttgcc ctctctcctg agtggaagca 69660
tcctgaggcc atcaccagat gcccagtctt ccagccaaca gaaccatagc caaataaacc 69720
ccttttcttt ataaactgcc ccacctcagg tattccttta tagcaacaaa aatggactaa 69780
gacaccagct gtctaggtat cccttaatcc agtcaagtta acatgtaaaa ttgaccattg 69840
caatattttt cttcttcacc aaagtttatg ggctatttta cttttgcttg ctttgtgaaa 69900
tgctaccttc atggcccagg gtttttctca acagtctaga gtttatctgg gaactctgtt 69960
tctgaatctc tgaggagtct catcaaagtc tctttttctc caattccatt ccaattcaac 70020
tcaaaaaaca ttgtaacacc tactctactc actggaacca tgcaaggact ttcatggata 70080
tagctgagcc ctcacctcaa gttgcttaca gtctgtacag acaggcagct cttccagtcc 70140
ccaccttcca ggaattagag cctgaaaaga gggcacaagg ggcttgggaa tctcaaagag 70200
gagggagaaa acaataataa cattggaata agaaaatttc tatttcaatg atcctcttgt 70260
tttcaaaggc atatcgtatt taaacccttc attcctatta cttattaact cagtacagaa 70320
gcagctgaag ttttatatat tctttttttt atttaaactt tctttattac aacttaagcc 70380
agagaattaa gattctcatc ctagagacct taaataaaag accatgtttt atctacttaa 70440
acaactccct ccaaccatat ctggtccagt ttttaaattg agaaaaacta cgtgaaataa 70500
cattccaaaa tgttgggaat gtttcttttg tgggcctctg gtgcacatag gcccccgcac 70560
tccacgcatt tcctaggagt acagtacttg ccatcctttc tgctttgaag cttcttctcc 70620
cagggccctc ccactcacag cttcccagag gcgtttcctg agaccacaca cccagtgggt 70680
ctcagagagt ttcctgtttt gtctccttgg cagcacttac aattgatgcc tgaaaatatt 70740
tacttcttat gttcttcttt ctctcctcca ccagcatgta aattccaaag ggctgggact 70800
gtcgtcttgt tcaacattgc ttaatcgccc atggcagagt gggcatgggt attgtaaaag 70860
acagtggagt tcaagcagca accagcatag tctgacttgc agccatctgt gtcattggct 70920
agttaatcat gaagtgaaat agataggaag cctactcaat tcttacttga tctgtataag 70980
tagaaaactt atatgactag tgaatgaaaa tctaacttga gtcatgtccc ctccatcatt 71040
cccagactgg agctgcttta cagacccaga accaactgaa tgaaggggag gctgggcctc 71100
cttgaagaag gatctttgta cccagctaaa tatttatact gttagtcttt ctcccagctt 71160
ccccaaaggg actgaaggcc ttttaccagg atgagtgtgc atgggagaaa ggaaataatc 71220
agacttctca gggcttgtgg acactggctc taaactgaca ctaattccag gagacacaaa 71280
atgtcactgt ggtcaaccag tcagagcagg ggcttatgga aggcaggtaa tcaatggtgt 71340
tttagctcag gtctgtttca cagtgggccc agtgagtcag gcactgaatc catcccgtgc 71400
ctgattccac agttctggac tgcataattg gaacagacct actcagcagc tggcagaatt 71460
cccacattgg tacagaatcc ccacatttaa ttggtacagg gcctgtccat cagctccatt 71520
tactaggaat tttagaacaa agagaagttc tagttcagag agtccaaact tttcattttg 71580
cagattgagg tgcagtggtg gaaccaggaa taactcagct aggacttaaa ttgcaattaa 71640
agtcatttta aacttgcaac tctgtaaata agtaaaatgt acattctgaa tccaaggttt 71700
acctgtctgt gtgttgattt tatcaaaaat gctacagatt tccagttgct gtgtcttatt 71760
tcattgctga agcatcatta gcagcactgt ctctgcaaat aaaggggact ggaaggagtt 71820
cttttttatt ttattttttg ttttattttg ttttttgaga tgggatctca ctctgtcacc 71880
caggctggaa tgcagtggtg cgatcatggc tcactgcagc cttgacctcc tgggctcaag 71940
tgatcctcct acctcagcat cctgagtagc tgggaccaca gatgcatgcc accacaccca 72000
gctaactttt aaaaatgttt tgtacaaaca aggtctccct atgttgccaa ggctggtttc 72060
aaactcctgg attcaagtaa tcctcctagc cttggctttc caaagtgctg ggattgcagg 72120
catgagctgc tgcacctggc caagctcttg ttttaaactg agatctactt gggtttatat 72180
ttttattgtc tccaaatgtg ctttcctctt aattcatcaa accaagtcat gagcatatct 72240
tcttagaaac acaaatatgt ccagctgaca tattgtatat gactttaaat gtttattcgg 72300
23
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
tacaatgaat gattttcctg cctggggaca acagcgttgt gtgggaagga ggactaggac 72360
atcaccaatt actcatctgt ctatcacttt cctaaacgaa tctcacacca agaaacagac 72420
actttcctcc cctcttttgt tatagaaaat tagaaagatg ggaaatggag ttggcaccat 72480
tgattcattt tcatttgacc cagaattaaa ttatttgtag ccagtcactg ttaattatgg 72540
gaatgatagt actttggatg ctaggcaata tcagagtcgg gagcttctca ctgagggcct 72600
ggcactgcta agcaggaagg tcaggagaaa aatataatag caacagaaat taaaatgaaa 72660
tttcaacgtg tctgtatatc gtaaatgaaa gaatagtaat ttcttcctct gtgaatagag 72720
cctaccaaat gtgtacttca tagctatttg caatattcat tcattagctg tattgagtta 72780
tagtaagaat accttggctt tgtagaaatc atcaccatta actggaatta ttcagaatgc 72840
tttatatgta gcatcccatt tatttccaca attgttttaa ataatttgtc atgtaaaaat 72900
tgtgtgcctt ttttattaga aaattgaggc acagaggccg ggtacggtgg ctcatgcctg 72960
taatcccagc actttgggag gctgaggtgg gcggatcacg aggtcaggag atcgagacca 73020
tcctggctaa catggtgaaa ccccgtctct actaaaaata caaaaaatta gccaggcgtg 73080
gtggcaggcg cctgtagtcc cagccactcg ggaggctgag gcaggagaat ggcgaacctg 73140
ggaggcggag cttgcagtga gctgagctcg cgccactgca ctccagcctg ggcgacagag 73200
cgagactcca tctcaaaaaa aaaaaaaaaa aaaagaaaga aaaagaaaat tgaggcacgg 73260
attgattatt gcttccaaat attcaacaga cacccagtct tctgaatttt ggtctgattc 73320
tgagctatga ggtttttccc ttcaaagtag tctttttctt aaaataatca tattggaaaa 73380
atgcagtcat ttaaaattgt ttaagacttt taaaatgtat tttatcaaac acctctatag 73440
tgcttgccgc gtgagagatg ctgtgttcag cactttatat tagtaaatat tgactctcat 73500
cagtgacccg atgagaaaga tgctattcat tattcatata tcttaactgt atgtcataaa 73560
tgaggaagtt gaagtacaga gaggttaagt tcctatctga ggtcacacaa taagtaatgt 73620
aatcaggttt catacccagg cagtctggct ccagggccca tgctcataac catcaggctg 73680
tgctgtgctt cagactcaac cagaaatgtc tctgatttca tatggcacaa cgtatacatg 73740
atatgtgctc atacgtgttt gaacagatca gtccaaaaat atctctggcc ccagagcatc 73800
agaagggaaa gaccttcccc aatatgtaaa atgaggcaac agtcaagatt atctttaatc 73860
caaaaataat ataaaaccca aattgtaaac agtgtgtcag tgttttctta gagctgcgtt 73920
gtccaacatg gtaaccacta gcttcatgta gctaccaagc acttgaaatg tgactagtcc 73980
aagttgagat gtgttgtagg tataaaatac atagcagtgg atttcaaaga cctagaacat 74040
ataaaagaat gcaaaatatc tcaataattt tacattgatt acacgttaaa ataatatttt 74100
acatatattg ggttaaatga ctatatatta aaattaactt tacgtgattg ttttgtttta 74160
atgtctcttc tggaaaaatt taaaattaca tacgtggctc acattttcta ttggaaagca 74220
ctgtcttagg caatgctttt tactttccaa caaaaaggct gaccttttgg acacattttg 74280
tttcccagta taccctttag caaaggtact attgatcaca aaggagaagg aacagaacaa 74340
aagatcttcc agacccacag tgtataattg tgaaaaatta gccatatgca ctctttagaa 74400
ttcactctct ggagcccaca gcaggaattt gtgtgcttca tgcctgcacc tccaacaact 74460
ggtgatgtgc ctggcccaga gtttgattgg gtgagggggt ggatagaggg ataatgggtg 74520
gataatgggt aaatgaaggg tgggtgggag gatgaagtgc aggaatggat gaggaaatgt 74580
atacagcaag aggctagata catctgcatg ttctatcatt aattatagaa catcataatg 74640
atcatcacca tctttattca tggcaccttt agatgctatg ggaaggcatt cttccaactc 74700
tatcaatagc taattttttt tcaagacagt attaaagtct tgatccatga ctaccccact 74760
tccctcagtg gtactaagtc tgctactgtg gatgcataga cctccacagt tttagggggt 74820
taggccacca atctgatagc aaaccagggc taagggagtt gaattgcagc tgttttcttg 74880
gcaatgcagt tttccccaca aattatatac ttaaagatca atatttttta aataaccaaa 74940
ttttactact cattattgct attcatagtt tacataatat ttagaaaatg acaacttcat 75000
atatcgaaga atattatttt tcaaattgtg gctttggaaa ttgaagacta aagctaccca 75060
aatcatccat cctttggcat tgccactggg caaaggaaat tcttgttctt gatttgatga 75120
tgagttaggt taccagtatg tgtttttctc ccatttctat ctttatcact gatagcaaac 75180
atgtgccttc tgatctatat agcaacctta ttttctgtgc caacctagcc tattcagaaa 75240
gagtgttagg tgtgtgctct ttcatctaag cctctgagca atctccccaa ggtctgtgat 75300
actctattta ctttcataaa gtggaagaga ctgaaaggtt catctttgga ctgctgctct 75360
aatcccttat ttctgggtcc aatttataac ataataaatt ggctctgaat tgctttgcaa 75420
tttgcctgac agttcagtgt cttctctcct ttttgttgat tccgagtaag aattaaaaga 75480
atatttttca tgcaagttat ttgtaattgt gattcctgac ataggccaga ccattcacta 75540
atctttttat aatcctataa tacttgtctt ataaaagctg ccagagtcag ctcttggtag 75600
actctgccct ctgtttatta tttgtattga acatgtgact ttacaatgat tattcaactt 75660
tctccttaat tatgttcttg cagaatgtcc catcttccag tggaaatggg aaatggaaga 75720
ttgaggtaaa gggctgcttg acaattatat ggtgaaataa gttagaatga attattcttt 75780
gttacatact agctagaaaa ttaaagcaga atagaattac tcagtagaat cccttctcaa 75840
gacaaagaat catcaaacat tccataattt ctctctaaaa tgccatgctt caagccattt 75900
24
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
taagtacaat tactcttttt atctataacc tccctctttc attgaagaaa cctaaacccc 75960
tccccacctt tgcctgaaac agcgtacatc agtgtgatga agcacagcaa caaacataat 76020
ccccactggg ttgccagctg aactggagta agacgaggcg tggctgtgaa attcacagtc 76080
aggcttccca gtttcccaat gaggtttgtg catacaggaa gacttggcat actctgtcat 76140
cctgttgtgc taaactggtt taacttgtgt gcctcacatt tcccagcaga ggagaaattt 76200
ggtctgtaat gctaacacat tctttgcctt gcacaatatg ccacttgtga atttgcctta 76260
gattattggg gaaaataaat gaatatattt catgatattt accagatgcc tagtattgag 76320
tcatgtgcag acctattgag gtgttgtgcg tgtgtgtgtg tgtgtgtgta tgtgtgtgtg 76380
tgtctgtgtc tgcttgcctg tgcattgggg agattctgat tcattctaga aatgtgttta 76440
atacagaatg ttttagtttc tttatatttg tataattaaa gttgcaaaaa cctagcacta 76500
atatttctta aggaataact ccaactctga aattccaaat attaacattc tatttcattt 76560
gtataaatta agttattttg aatctggcta atctcacacc tacgtagagt gtgaatttct 76620
attattattc cttctgtgcc tcttattacc aaggataatt aagatttgac atttttgaca 76680
gctaataaaa tagacttaat atccaatact gagaaaaatt ctttataagc cactctgatc 76740
tcattgcttc atttatgtaa atgttccatt aaatataact tgaaaaaaat gaacttgaac 76800
agatttcaac agtagctaac tgatcattct ataacgaaca ctaaggtctc cacgtaaact 76860
ggatttaagg atgtgttgcc catcagttaa tgtaaaagat atttgtgtct tatattatgg 76920
acttttatat aaaaccaatt atattttcca cattagcccc ccactgcaaa actttcccaa 76980
ccctagcgct attaatattt tggactggat agttcttttt tgtaaggagc tgttctgtga 77040
gttgtagggt gtttagcagc gtctctggcc tctaccctct agatgtcagc agcagccccc 77100
aaggtatgac aacctaaagg tctccagaca ttgccagata tcttcaggga gcaaagagga 77160
gcaaaatcag ctctatttaa gacctactga tacatgggaa tttaaggact ttcatttaca 77220
tttttttaat ttcttcatat attttcagaa attgtatctt agtgtaataa acaaacaaaa 77280
aacaacaaat aaggaaattt ttaacccctc acatgcaata ttgaggtaga atcataatgg 77340
gaaacttctt tttaatttat aagatttggg ggttggggag tggcagattt ttcttatctt 77400
caaaacttac cagaggaacc aaattatgtt gtaagacctg aaagcccttg ctttatctct 77460
tccctaaaca tttttctgat gatcattata gcatctgcaa cctttttctg aaatgctcac 77520
aggtgttata gaagctcctg gaatgcccag tgggattgcc cagtgtcccc tgagcagttc 77580
caagggagct ctctccacaa caaggaccgt caggtcattt ggtcaagggg tgaccacagc 77640
aactaaggca tagagcaact caggaaccct ttgggggctc cattgctgtc aaaactttca 77700
gtctagcacc ctcaacttct attatggtca tattggttca gaaagtgagg tgtatctttc 77760
tattcactgg agatctttaa atcatcagaa cagattactc caggagactt ggtatgccaa 77820
ctacaacatt gaagagtctg ggaaattcag ttcttttgcg taaacactta gcacatgtcc 77880
actgggccag ttaatgggga ctcaagaatg aatatgataa agtccctacc ttcaaggggt 77940
ttatggggtg tgaaacacta gtaatagtaa tagtttcttc tctgtttaaa ggacatcaca 78000
gtaggacaag gagtaatttt ttccccaaaa tatatcatca gcccacaata agaaatgtga 78060
atgcttggat tgttttatat atcgctgagt ctaaataact ttgcaaagga tttaaataga 78120
agccttctct aattcttcag actcacttat cccttagtat tccttagcac ttccttgcag 78180
cctgctttac aaacaagtat tgagagacat ccagaaaaaa taaccacgtg aaaaagaggc 78240
attgaataaa aaccagccag attgcctaga agcccctaca agttggtgtt ttcttatgta 78300
acaatgattt gttgaaaact aacaggcaaa tctacagtct tccacttgaa attatagcca 78360
cactgcaaaa aaaaaaaaag aaagaaagaa attatagcca caccgaagta cctttatctt 78420
ggtccagagg gtctggcata tcctgacata caatagtgcc ttcatgaagc ttaagttata 78480
gctattaaga ggatgtccag agtcctgatg aaacattaat gaggaagtga aaagtcagta 78540
aaaactagga ctatttttat gcattgagaa gttaagctgg gattttatgg tgcagagaaa 78600
cggattcttc acatcaaatg aagaaaagga gtagtattaa attatttgtg acacagatat 78660
tttatttcat actaggattg tgtgttataa tgaaaccact tataatctgt tcaaaatatt 78720
gttgagtttt aaaattcaac tagcttacta taccaataag catcacacat ttgttttttg 78780
tcatggttaa tatgtgatat ttgttttcag ggcttttaat gatcacagat agcaagcaag 78840
catgcagctt accccttgaa agggcctcac tctgtcaccc aggctggagt gcagtggccc 78900
tttaaggctc actacagcct caaactcctg gattcaagtg atcttcagcc tcccagtggt 78960
ctttgtagac agcctggtgg agtctcatgg cacagaagat taattaaatg atgtctttca 79020
attttactat ctgcattcca tcaaaaaata aaaataaaat aaaataaaat tcaactagct 79080
atccattgta tgagctcctc aagaagccca ggcaagagaa ttgtatgagt ctaatacggt 79140
gcagtataga gccagctctg agcccccacc acatgcattt gtttcttgtt tgagggtttg 79200
gttatggact tggacttggt tggctgcctc tgtgtgtgat ggtgctaaat ctgaagacag 79260
gttgagtcac acctctaaga attttaggta gaaccactgc tgtcatagcc cagagaaact 79320
gatatagatg cctaaagttt gtgaatttta aactctttga acaggaactc ccactaagaa 79380
acacactgtg acccattata tattttatat atgtatatta tacattacaa atacatttat 79440
aacacaaatt tcatgaaatg atacttatgc ttatccttat tacttaggtt gcacttaaat 79500
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
atttttatca ctatctcatt ttttaaaatg ctctttaccg tttatagcta ctaaactaaa 79560
ttttgcaacc catagattga aaagcaccag cctaagtggt cttcccatgt ccactgatgc 79620
gtaactatgg catcacttta aacagaggca aatacatgaa atggtgccat attttcacct 79680
accagttgat gcaggatgac attgtaaatc ccgtggttta tttggtacac tagtttgctg 79740
ccaaaatgac ctttccctaa attgcatttg ttaagaacac tgccaatact tctttatcat 79800
atcaaaggct ctaagaaaat cagttattgc taaataaata aacatgcttt agtccttgac 79860
atcaaatttg catgaaatgt caatagcaat ttaccttaca gaaatctgaa caaaaatgaa 79920
tcagtgcaca cttcctaact ctattagatt ttgtgtcatc tcgacaatgt acttatgttg 79980
tgtgtctttt taaacaaata ctttaaaatt cacatattgc ttggagtgat agtaatattt 80040
ttctttacta ccttgctttt tgaggcagtg gttcctaaag ttttagggcc aagacctatt 80100
tgaaattctg atgaaatcta tggcccatct cctaaaaatg agcactattt tgattccata 80160
aaccctcatt tgagcaccct taccctgggt agaggtcact gtttctgtca tctccccaga 80220
gctcagtcag cagcttgtat aactagctcc tactgaaagg aagcattata ttccattcta 80280
ttgatatcta aaccaccagc tccttatttt attctacaag tgtccctttc atgtttactt 80340
atttagcagt aatatctact ggattgggac tagagctaat cttggaaatg accccgtgag 80400
tcatttcaac taatgtttgt caacacatta acctaattta gtgctgctgt ggggtagcag 80460
tgagtaaata cagattgcag acagtaccct agaggtgttt gaagctgttt ccaaagtccc 80520
cacatcagga atgggacatg gcacagcatc agtttttaaa aacaagttaa ttaaaatatt 80580
cactagatta taggaaatca atgtaagttt tgataaatat cacacaaaac acaaccatga 80640
gctgaaaagc caacggaatc tcatgttaag ctctatcaac cacctgagac aaacaccact 80700
gccacctctt ggtaacatag cttaattgca ggttcaactt tttggaggca taataaacct 80760
atattaaata cagagactat aatttttaca ttgtaaaata atagaaaata tatgtttgtt 80820
gatatgctgg gatagggaga aagcatgtgt tgttagagtt tgctttggct ctgccctttg 80880
cgtctcactg gagctgtcca caccctgatt ttcatgtatt ctctgtacct cttggcagga 80940
ttggaggagc catacccact gtgttctcgt actttgctga agtcctggcc cgggaaaagc 81000
ggggcgaaca cttgagctgg ctctgcatgt tctggatgat cggtggcatc tacgcctctg 81060
ccatggcctg ggccatcatc ccgcactacg gtaagaggct ggccttgccc cagctgggca 81120
gtcacttcat tttgcttcag gctccattcc catcttcttc ctctcttcta agggccactt 81180
tgagaaaaac tactcatcat catgtttctc cctttcatag tgtccaggag tttttcccct 81240
ctgtgtagag atctgtataa agtaagactg tatcaaatgc ccttctggtt atatgaatgt 81300
aatgggggga aaatattttc ctctgctatg ctttcacaag atttctggtc accaaaatgt 81360
gtggggtgtt ttctcccact gaccaattct ccaacagctg ggtgtgctac aatttaactc 81420
aattctgaca ctatgttact tggggttgga gtcaaatggc acagattaag ggctcagtcc 81480
cacaagactg ccccccattt cagaggccaa ttgtaagtcc aggcctctgg aacttctgac 81540
tgaccagcta taaatgggag gttcctatac cccccaccct caggtttgat catttgctag 81600
aatggctcac agaactcagg gaaacaactc tcattcacca gtttatggca aaggatatgg 81660
caaaggatgt agatgaacag ccaggtgaag agaaacacag ggcaaggtat gtgggaagag 81720
gtgaggggct ttcatgagtt ctccgcctca ccaccatccc agtgcctcca catgttcaac 81780
aacccaaaag ctctccaaac tccatagttc agggattttt atagaggttt ctataggcat 81840
gattgattag gtatgattga ttattaactc aatccccagc ccctctcccc ttcctggagg 81900
atgggggtgg agctgaaagc tccaagctta tgatcatggc ttggtctttc tggtgaccag 81960
tcccactcag gagcccacca acagttgtct cgttagaa~ta aaagatgctc ctattaccca 82020
ggaaattccc aggaaataga agtgtcagga accatgaatg acgacgaaca catatgtttc 82080
ttattataat cacagcatca tagtgtacta ttgcatttta ctgtgtttaa agtaatctct 82140
ttcatttgtt tcctctgtgc tatcaatgct ggggattcct ctcccctctg cctctcctac 82200
cctaagtaga ggtgggagga tacacagatc agcagcagca tataagacaa catgacttat 82260
agttgttctt gggactgcca tacatatcaa acacacagaa gcacctttac tcaactcttg 82320
tgggctcagt ccaaagctca gctccacatc agccccacct cttctggcaa ctcagaactg 82380
gaatgggagc aggaaaactc ctgttgtatg gcctgttgaa ctgcagaact ttagaggtga 82440
ctccacctgt ccacactccc agatcccctc ctcagtttct cctaagctct tactgtttct 82500
gctgcctcag aaacctgtac atgttgcatg tttctgagca atacccctca gaagccaaaa 82560
gctccaagtt gcttcatcta taagcagtga aatgcatgga aaggtcaatt gaatccagaa 82620
atcaaattca atatatcagg aaaggggaga gaagtgttct ggttgcgtgt gtgtgtgtgt 82680
gtgtgtgtgt gtgtgtgtac atgcacacac acgtgcacat gtgcccaaac aggctaaagt 82740
ttgcaaaata ggaataccgt ataaattaaa accacatgac tgtcagtaaa ctcactctct 82800
gctctaatct aaaaaattag tatttcaaaa actcattctt tgctcatttt ttatttctct 82860
gcttttgttt tagaacatgg taatatttca ttataaaaca cagttctgtt agggaacaac 82920
cttggtgaaa ttgactcatt gctaaaatca gacatagact ttagtattta gatgtcttta 82980
acttgttttt ttaactgagt tttaacatgc tgagttacaa aatcatgtca cacgctttct 83040
tttgatgaac ctgaaaagtt acatgaaaac tgaagtcgta tttaacaaca acaaaagcaa 83100
26
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
gatataagga attggttact tggcctgaag agaacttctt tgctgaacat ttacattgag 83160
ttcagggtgc caagctcaag tggtgggttt ttcattttta aactcttagg tcacttatgg 83220
gccagagatg ccctcataca ggcaggtccc catatccacc atgagacttg tcttctgcca 83280
ccatcCaagg aattcaagat tttttttttt taatttgaga tctcaggacc tcagccaccc 83340
tcagaacctt cctcggtaga gcagcatcta actcatgccc cactcaactc actggttcaa 83400
gtggaaaagc cactcaggaa ttgctttata ggaactatgt ttaagaaggt gaataggaaa 83460
tgcagagtcc agctttaccc atcattgctc cagagccact gctagtctat ccattcatac 83520
tctttgtgca tattcaaata agggctgctc caggtaggta gccagaagtg ctgccccact 83580
tatcatctct gccaactact cacagagggc tcaacctggc aaatttatac agcagctgaa 83640
gtgctttttt ttctctctct ctctcttttt cttttttttt ttttaatacc ttcttagccg 83700
gccttcaggc cagccttgtt ctcttttcag cctctgatga gagccttgct ccttggctct 83760
cgctgagctt gctctctgga gccccttgtc tccctggtta atggctgtct ctttcctggt 83820
cctcccttcc catcatatcg ctcagaattt cacctgctgg ccttcttaaa cacgtaactc 83880
acactggctt ctgcagccta cgtgtgtccg tgtgtccacc aagcccgggc atgactgttt 83940
ttatttttag ttttttttct cactaggaat tcagttcctc tgactctgcc tttcacacat 84000
ccctgccctt gaactctgga tggttccaga ttttctctcc tgccttgtca ccggtcagtc 84060
agtggccaac ccacaaagaa taagcatgat caccactttc ctttttccaa agtacactta 84120
agatacttct tgtttatagc tacaagggct aattaattca tatataacta gaatgaggga 84180
aagttgggaa tcataaacaa tgcatgaaaa gttacaaaaa acaatgacga cacaacaaaa 84240
ttaaatgtcg gaggctgaca tcaactttta tgttgacaac accttgagaa ataatgacca 84300
ggttacgaaa tgagtaatgt caatatgaaa accttatgcc catgctttac taaaggcttt 84360
ccatggccag aagtgtggca aaaaaaacat atatttcatt caattctgtc tgcaaattaa 84420
aattaaaaat gaaaagcttt aaaaataact ttgctgaggt cattgttgta ctcttgcaat 84480
ttgaagagtt ttaatttttg ttttaaagtt taatttttaa aatcctgtgc tgaggcttct 84540
ggagtgctaa tgacattcta tatcttgagc tgggtggagg tttttaatga atttttgaat 84600
tgtaaaattt gttaaactga gtgcttaata tttgtgcatg ttactatatg aatgttacac 84660
tttttcacaa acacttttta gtaagtttac cacctccccc ataccctgaa aacactcatg 84720
cttggattta ccccacatca atcaaatcag attctctgga caagagacac agcattgaga 84780
gtttttattc ttattttttt cagctttatt gaggtatgac tgacaagtaa aaattatata 84840
catttaatat gtacaacttg aaggtttgat atatgtgtac attgtgaaat gattaccaca 84900
atcgagctaa ttaacatctt tcatgtgtgt atgatgagaa cacttactct cagcaagttt 84960
caagtataca gtacagtaca atattattaa ctatagttac catgctgaat attagatctc 85020
cagaacttat catcttataa ccaaaagttt gtgccctttg accaacatct ccccccatcc 85080
ctcaattcct ggtaaccact cttaactact ctctctttct atgagttcaa cttttcagat 85140
ttcacatatc ggtgagtttg tgcaatattt gtcttttcat gtctggccta tttcacttaa 85200
cataatgcct tccaggttca ttcatgttgt tacaaaggga aggatttctt tctttttaaa 85260
ggctgaacaa aattccaata tgtgcatata tgtgtgtgtg tatgtatgta cataagtata 85320
cacatacagt tttaaagctt tactgttttt acagttttat agttctgaac ttacatttaa 85380
gtctttaatc cattttaagt tgatttttat ctgtggtgca aaaggaggtt ccaatttcat 85440
tcttttgcat atgaatatcc agctttccca acaccattta ttgaagagac tatcctttcc 85500
ttatggggta ctcttggtgc ctttatcaaa gatcagttga ctgtataagc atgggtttat 85560
ttctgggctc tctattctgt tccgttggtc tacatatctg tgtttatgcc agtactatgc 85620
tatttcaatt actatagctt tgtaatataa tttgaaatca ggaagtctcc agctttcttc 85680
ttcttgctca aggttacttt agctactcag ggtcttttgt tgttccatac agattttagg 85740
attgtttctt ctatttctgt gagaaataac attggaactt tgatagggat tacattgact 85800
ctgtagatct gtttgagtag cattgacatt ttgacaatat aattctttca atccatgaat 85860
acaatatatc ttgccattta tttgccatct tccatttctt tcatcagagt tttatagttt 85920
tcaacataca aatcttttac cttctgtgtt aaatttattc ctagtttatt ctttttgatg 85980
ctattgtaag taggattgtt tcaataattt cttttccaat acttcattat tactgtgtag 86040
aaacacaact aatttttgtc tcttgatttt gtatgctgtc acgttgctga actcatgtat 86100
tctaacattt tttggtgaca tctttagaat tttctatatg taagatcatg tcacctgcag 86160
aaacatactc ttgttttact tctttctttc tgattttgat tccctttctt tctttttttg 86220
cctaattgct ctggctagga ctttcagtac tatcctgaat agaaatgaca agagtcttcc 86280
tagtcttaga gggaaagctt ttagattttc accatcgagt atgttagctg ggagcttgtc 86340
attgatggcc ttgattatgt taaggtacat caatagtttt taaagcttct gggtggttct 86400
aatgtgcagc caggtttgag attcttcagc ttcctgtcca tgaaaggccc aatgggtgac 86460
ctcagaagtt tgtttttctg ctgggaccct agaacttgat ggtatgtatt gggtgggagt 86520
ggggggatgt gaatgagaat aaggagaaca gtgagcctga tctgttcagt gtttgagttt 86580
ccgtattgtt ggtttcatcc acgcaaaaac actttcctgt ccatctggtt tagaatcaca 86640
cttaaaaagt aactttgaca tctagatatc tcttcttttc agtgatgcat cttttaactt 86700
27
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
tacctaaaat agacaagggg ctaaaagtct atccactttc cctgaagcct ctgaaaaaac 86760
taaaggctta tatctctcct ctgaaatctg cgttgtataa atgatgccac agtccacaga 86820
aacaagagcg ggtccgcccc tgctgctctg tcctctttca ctgaggaggc ctttgagtgc 86880
ccaccactcc ctgtgcattg agtttgcacc ctgcagagaa tgaacacagg agtgtaataa 86940
agtgactcag agccaggacc tcagccttct catattagta atgggctttg ctctggagag 87000
aaggaatgga taattggcat ccacattgca agtgtatttc aaatgataaa taacacagag 87060
gctcgtggac aaaggtgttc gctgtttctg ggtgtcgccg tttagtaatt agggcagagc 87120
agtcagttca caaagcctga gcctgatgct gtgagactgg aattctggcc cctgacaagc 87180
ctctgatgca aagtagacct tgagtgggtc actaaggatt ctggaaggaa atctggaagg 87240
attcttgtga agttagacac tggaataagg gattggattg ctctaagggc tagtgagtta 87300
agacatgggc tgtggccagg agtttttggt ggacagtgga gtccactcag cctgctgaga 87360
cctagctttt gtaccagtgt ggaataatga tggtgctttc tcttcatatc actgctatca 87420
gagttgaaat tttccccgga gacagatggt tgctaacatc tgaaggttcc caacatgaat 87480
ggacctcacc caatgctgac aagaccttag gaggcagagt gcttggctgt ggtcacacca 87540
gcaccttccc tgccaccact ccattcagga acttctgggc catctgtgtc tcccccatgg 87600
caaagagcag tccactagaa caagggagga ggaagagcaa caaactgaga aggaaatgcc 87660
cttgcctcaa aaatcagtgc ctgtaagagc tgaaaatagg tatttagagt tggcttggcc 87720
gggtgcagtg cctcacacct gtaaacccag tgctttggga ggctgaggca ggagaattgc 87780
ctaaagccag gagttcgaga aaatttggct cttcttgacc taacacccgt tctcaagcct 87840
aggccttctg acccatagtg ggaattggct actggttcag aaataaataa gcagtatctt 87900
caagaacaga taggtcaggt gtaaaggttc tctttcagcc ttcctgccct tcctttctct 87960
ctccacactc tgtaataatc aggacacatc agcatttaat ggaagcccca gtacctcagg 88020
ggaaatggga acagagagct gaatcacagg cactttctat gttggctgag agctttcatt 88080
aggcccaggc tttggaattg ctgctagact gcggcactgt ccgctccatt aaaccctctg 88140
tcatcatttt cccttttaaa ttcatgtgcg catcagatga ggtgttctcc cctgctgtgt 88200
gcattacagt gtgtagcaaa ggtgttcaat tagctaaatg ggaaagttct tgttttagcc 88260
cgaaaggcca cagctctgaa agcccacagc acactgggcc tcatctagcc cccagaatag 88320
ccccatggcc tctatctggc tagccctgca ctgcagatgt tatagggtca aacttggggc 88380
tcccataaac gtcagactta ccccacccta cgtctccagc catgatgcga tgtccaggct 88440
taaaaataca tatattccca aggttgtcca aattaccaaa acatccctac ttcctttggc 88500
tggcaggaga ggcaatagct atctaattga agaaggaagc agggaaggat tcctgattcc 88560
tcaaccagac agatattcaa caagaaggaa gaagaaagtc agcttccttg ttcacaagaa 88620
ggaatgcact gtggccaagg gaaagaggta taaagtgaga ggtattttga gtctcagacc 88680
tgctacttct gtgagctctg tcaaagcttt tcagtctgat ccagcccact ctcctagtca 88740
gcattgtagt aaggattaag caaaataatc ttgttagaat acccagtgtg tatttgatgc 88800
tcaaaaatgg tagctcttgt tgctatcagg tcatcctaca tgggataccc agattgccag 88860
agcagcccaa atgtgagcta catgaaaaaa gagggagaag gaactaaaaa gcataaaccc 88920
tgcctaaagt ttgagattag agaagtcaga gaaaagaaaa aaaaaaggat agaagctaag 88980
ataaaagaga accaaggtat taaaacaata ctgtaaaatc ttgatgactt tttacccaat 89040
aaatcagaat cctccatttt tctgttgggt aatttaatat tcaaggggca tgtactatat 89100
aaagacccat tttccaaggc catggtgcta tcgttcccag atagaaataa cgagagtctg 89160
acaactctaa cctgtgagca tcaactacca gctgattctg cgggttatat gggcagcacc 89220
agagccaagc tctatcgcag caccaagatg tcagagatcc tggagacagg cagtcagtgc 89280
aattgttaac agcatgggct tagagggaag gccctacttc caagccacct ctgccctctc 89340
tagctttgtc atctttagga cagtaaccta ggctctctgg tcctctgttt cctcatctat 89400
aaaacaggga taataatagt atgacctaat acaattgctg tacgatcaca ttactgggta 89460
tatacccaaa ggattataaa tcattctacc ataaagacac atgcacatgt atgtttattg 89520
cagcactatt tataatagca aagacttgga acacacccaa atgcccatca atggtagact 89580
agataaagaa aatgtggcat atatacaaca tggaatacta tgcagccata aaaaataatg 89640
agctcatgtc ctttgcaggg gcatggatga agctagaaac catcattctc agcatactaa 89700
cacaggaaca gaaaaccaaa taccacatgt tctcactcat aagtgggagt tgaacaatga 89760
gaacacatgg acacagggag gggaacatca cacaccaagg cctgttggtt gagggaggca 89820
aggggaggga gagcattagg acctaatgca tgtggggctt taaacctaga tgatgggttg 89880
ataggtgcag caaatcacca tggcatatgt ataactatat aacaaacctg tacattccac 89940
acacatatcc cagaactcaa agtaaaataa aataaaataa tacaattgct gtattaaagg 90000
atggcataca aaaagtgcct gacacagtgc ctggcacaaa tgttcattaa gtgtagtctt 90060
tgcacagtgc ctggcacaaa tgttcattaa gtgtagtctc gtattttcat atttgggact 90120
gaagtttggg tcagggagga agaattaaaa agaggaatat ttaaaagatt gtatttacct 90180
tttcctaacc atgttttctc aatggttcac ctgtcatttg aagttgtgga gatggataag 90240
aaagattaga agtacaggaa cagaacttgt aaggcatgcc aaagtgtcct gtgcccacta 90300
28
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ctctagggtg tcagggcagg cctcacatga taactggggt tcctctcagc agcccagcca 90360
ggacaggcag gtgacaggtg gcccagagcc ctgtcctctc atccttgcca tatttcactg 90420
agtgttacag ccctgatgat tttatgctca agaggcagag taagacgtga cagagggaag 90480
tgatttggta cattgactta taccagtatt atttcttaat ttgttaattt aatttgttaa 90540
ttttacgatc aatcttatat tttatctcaa cgaaagaacc cttggcttcc gatttccttc 90600
agtggtggtt cacttttccc catgcaataa taagatgcat cagtcaaata aacattgatg 90660
atatagtctt taagggctta aaaagtagat gtgaaatttt aaaaaaatta tcatgtgttt 90720
aagtgcatgg ccatttcttt gaatgcatct gtttcattgt tcatggcacc cagaagcaag 90780
tgtttggaaa gtctcacgtg ctgaaacatg gccacaagcc aggtctccca gcccagcctc 90840
ttgtctctct ccccatcctc tttgtttagt aggggtaata tggagcctgt agggaaggca 90900
tctatcagct aggatgcaga tcgtccccac agacccaaaa gacgtctatt tactttcttg 90960
agagattgag agttctggcc agatctggga aacacttttc aacattctca cgctgacctt 91020
tgaggtatgt atagtgaact cagaggagct gcctccccta ctgtgaatta agccaattat 91080
tgtggaattg aagtctgaac tcttccattc aagctcacca gaaaaggaag tgatgtcagg 91140
catttactga gagctttctg tgtcccaggc attgtaccag gtaggttatc ttaggcacct 91200
gatcttaaaa tggggaaatg ggctttaaaa tatggcgatt tcctgctctg gtgctcatgt 91260
gaagaactgc atgttcctat gctgatagca ctgcaggtga ccctgtcaac ccggggtttg 91320
tgaccatatg tccctctggt aagtgggtga agcccacaga cctcttccca aaataatgtt 91380
tttatataca taaagcgaaa taaataggat tataaatgaa atcaattgtt tcaaaatagt 91440
attatcaaaa tatattagca tactaagtaa caacatctag tgacagggct agtagctacc 91500
acaattttga agtagataca agtgtagaag ctggaaatca actggtgcat tcttctcaat 91560
cctaggaaac cactggaccc cagattctct ccagaatcca tttcttcact ttagttaaaa 91620
attagaattg acaaccgggt gcagtggctc atgcctgtaa tcccagcact ttgggaggcc 91680
aaggtgggtg gatcacttga gctcaggagt tcgaggccag cctgggcaac atggcaaaac 91740
cccatctcta ctaaaaatac aaaaattagc tgggcatggt gtcaggtgcc tataattcca 91800
gctacttgtg aggccaaggc accagaattg cttgaacttc ccgggaaggg acactgtctt 91860
aaaaaaaaaa aaaaaaaaaa gaattgacta gatagaaagg aaaagccaaa ttaacaaagg 91920
tataagcaag atataagttt gttttatatc acatgaaagc agtctggata gaggcagtcc 91980
ggggctggtg tagtagctca gcggtcatca tcaaaggccc caattccttt tttcttactg 92040
ctcctccatt cgtagcatgt tccaactgta gggttggaat tctaggcagc acaaagtggg 92100
aaggattcat aaggatatgt ctgtactccc agaaagaatc ttcctggaag tcccacacaa 92160
atccacttga atctcattgg ccagaatttc aagacaaaga aa,gtcttgga aatgtagagt 92220
agctgtttat tcccggcagc agggagaata caaatcagtg tagagtggac acagggaggc 92280
aactggccat ctctgctgca ccatttcatt cttactctgg aaaatctttt aggaatgttc 92340
ctgcatttga taatctaatg tttaagtgtg gcccttccat ccatttttgg atgccagaaa 92400
cagaaagcaa ctctggctaa ctcaacagga atttcttgaa aggctattgg atagctcaag 92460
gaatggtcag gaaggctgga gaaacaggct cagaaaacag cagaagacaa aggaatcctt 92520
ccttagatgg cttacatggc ctttcagcaa actcaaaatt tcagcactca gacccaaaga 92580
aaacacttac agtttaacaa aaattggccc aaattacaca acttcagttt tagtgcccca 92640
aggggctttt aatattgtta gattttcttc caaagtcatg gggagtcgct gtctttgcaa 92700
gatgcaatcc ttaggggtgc cgcctgctgt tagtaaatca taattaccca agcatgacgt 92760
tgtggacaag tcaccccagt gccctcattc agataagtgc tgagtacaca ctaatgtgtg 92820
ttaggaggag aatctgggcc catttgaagg taggcatagt ttgggctgct gtggcctttc 92880
tatttggttt cagagaatgt agatacaggg gtctcgagac cctcaagcca gtgtgactcc 92940
tctttagcat ttcagagtgg attagacttc cctggtactc agatcacagt tggctagatc 93000
ctgggtcttc cccaaaccaa atgtgaatct gagccttacc agcagactag atccccctca 93060
aatcctggag accttgacat acagaagaga caaaggcacc catacctatg gtaaataaaa 93120
gtagtaacat tggttttact tattgtagat atagatgcct actccatgag ctgttgatgt 93180
tggggtttgt cctgaaaaca aattgtgaat aaatcgtgaa tacattttat agtattaaaa 93240
aattagtata aaaaattaat ggcaatatca aatgctggaa aggatacaga gaaacagcat 93300
ctctcatgca ttgcttgtgg aaatgaaaga tagttatgga cactctggaa agtggtttga 93360
cagtttcttt aaaaaccaaa catacaccta acatatcact cctgggtgtt tatcccagag 93420
aaataagaac ttgtatccac acaaaagcgt atgcatggtt atccatacaa ctttatttgt 93480
aacacccaaa agctgaaaat aaccaaaatg tcctgcaata ctataaaatg ctactcagca 93540
ataggaaaga agtgattgct ataacagcat gtatggacct caacagcatt ttgcagagtg 93600
aaaaggccaa tctcaaaagg catttatgta acattctaga aatgacaaat atagaggtgg 93660
gagaacatgt taattattgg cagaggtttg ggatgattgg ggggtggggg gtgtatgact 93720
ataaaggagt tgcatctggg agaactttgc gttgacgtca tagttgagtg tcttggtttt 93780
caatatggtt atgtgaatct acacatgtga taaaataaaa cagaactata tacacacatt 93840
gcactaacgt cagttccatg ctttcgatgt tgtctatagt tatgcaagat taaccattga 93900
29
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
gggaaactgg atgaagtgta catgggatct ctctgtacta tctttgcaat cttctgtgaa 93960
tctataatta attcaaaata aaaagtagaa aacaaatgag taataccttt caagagggca 94020
gtcttacata tattatctat ttattctaac aattttgggg ggaaaaacaa cagtaactat 94080
tactaccctt ttttgaatag agatcctgtt gcatagaaat ttggtcattc cccagattga 94140
cacagataat ggaagattag acatgagaag caatcatttg atccttattt cttcaaaatg 94200
ttctctgtac atgaagaagc taagaaactt ctccagagag caacacagag ttggaatggt 94260
gtctggggca tttcaactgc ctctacagtg tgcatgtgtg gcccagatgc cagctacccc 94320
ttgtctgtcc aggcgatgct ctcacagccc cttgccaagc attcagagac ctaagnnnnn 94380
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnngatat tacacacaca 94440
tatacataca tatcaaaaga catatacata tataaataac catagaagat ataacataat 94500
agcatacaag aaaattatta tagttatata tcaaaaatta tataaagaat agaagagagg 94560
aaagaatagc atagaagtac agagaagtag agaagaacaa gataagagcc aacattatat 94620
tataaaaaat aatattataa aaaattaact agaagcatag atgagcacac acatagtagc 94680
acatagatat ataggagaag atagaatata ggagaagaat agcagagcat aggagtatga 94740
ggattacaat gagtcatggt caggccaaat gcactacaag aatgggtgac agagcaagac 94800
catgtatctt tatatactat actatatata tatatatata tatatatata tatatatata 94860
tatatatata tatatatata tatgtatatg tatataaaga tttcataaga gactgaagaa 94920
gttttttaca tcacatgctt tataacccac atttgataaa tcctgatttg atacttgtgt 94980
ttccctcaaa gacaacataa cttctctctt ctcctcccaa cacagcatgc aacttctcta 95040
gctcctctct tcctccagga atcaactcca ctgtcacccc caacccctta ttactcagag 95100
aaatcaagct cttccagctt ctggctccaa acttgcatac ctgtcttaat ctgctcaggc 95160
tgccataaca agataccaca gactgggtgg cttaaacagc agaaatttat ttctcacagt 95220
tctggatgct tgaagttcaa gattaacatg tcggctgagt tggcttctgg ccaccttctt 95280
gctgtgtctg tacgtggtgg ggatggggaa gtggtggggg gatgggcatc tctctcttct 95340
gtttttataa ggccacagtc ctgtcagact agggtcccac ccttatgagc tcatttaacc 95400
ttaattacct cccaaagacc ctatcacttt caggcatatt gggggttaga gcttccacaa 95460
tgaatctgag agacataatt cagtccataa aaccacccat tatgatctta gcaaacaaag 95520
ggccctctag ctactagaag ggctccaccc tttgaaggca ctgggatttt ttgggggttt 95580
ttttcctgct gttttattgt tagctaccta ctgcaaacct aacttgaaac acggatatct 95640
tttctctcta aaataaggtc ttgtgctatc attatgattc cataagaacg ccccatctca 95700
ttttcctgca catttttata ttctgcacta gacaaaatat gtatgtgtgt gtatgtgtgt 95760
atttctttga aaataactgt tctgaaaata cccttgctgg gctgaattta tcattaacct 95820
aattagatcc tttatcaaag gcttacaatt aacaccatta gcaagtctta gtaactcacc 95880
gctgtgtttc taaattaact ttcacctgta gggacataaa aatcgccatc attttctaaa 95940
atattccttg tttgagtgtt gtaattgttc cccagatttc catgaataat attaagtgac 96000
ttacaggaca agagatactt acaattttgt ggaaagggag agaatgcttc cctcatatct 96060
tagggtacaa gttgaataac ctcaaaatcc tttttcttcc atatctagag tggggtcccc 96120
agaaggagta ctcccttaat ttgcccaggt tgttaaggta aaaaattgta gagctaccca 96180
tctgttcttc acaagcacag tattctttca tttgcaagga ctaagccaaa ctttattttt 96240
ttacataacc taccatcaga ggcattgatg ttaaattacc taccagttcc ctattctctt 96300
atgggctgat tttgtgtttc tattttattc agctgtctct gtcaaaatca agtttatatt 96360
tgccctgagt tgctttgaga aggtctcaag agaatccaag gggacactgg gtatggcatc 96420
ttagccagtg tggccattca gcaccactgg gccacactga tgtttcctat gtgactaatt 96480
tcctgagatg tacaatcaac tgcctagagt agagcattag aaaaatagat tgctaaatac 96540
gacagagtgt gatgttgaaa gcttgacagg gaatctaatt atgcagtgaa gtaattccta 96600
gggatccttc tcaatgatgg aaggggcttc ttctttggga ggtttctgat cttttatgta 96660
acactgaggc atgcttccac ttgaggataa attaaattgc atgatccact tcttcctcat 96720
tcccatgctc agccctcaag gagctcttca gctgtgctgt ctgtcacacc atggaatgtc 96780
taagacaaaa gggagttact tggggcttgg acaggatact tgcaaagagg ttggaaaaca 96840
atgagcctga gaactgggtg gcaggagatg gcagtggtca gccttgagat ctggtgaaat 96900
caaaggaggc agactgtgat gaaaggaaac agtatgggtt agaacagagt gaagtggtta 96960
ctgtagatct taatcaaaca aaaaaaattg ctcttgtgta atctagtggt agccgaagag 97020
atgagccatt taagccattt atgtattccg tgcccagtct ttccaatgat tagtagcaca 97080
tcttaatgag ataaccagca tgggttaggc aactggttga gggccatgga gcaaatatat 97140
tttctctgtc taaagtatcc agcaaagaag tggattcaat ggctttgcct ccattacagc 97200
taaacaaact gggattgagc cttcatttaa attacttgat gagagtataa cactacattt 97260
tgggtttggc aaatgctcat tgggcatccg tctgtgcctg gccccatgct ggaagccaga 97320
ggtgcttcta ggctaaggtt ggactgcgaa gaaatctctg gtctggggta acagcaaaga 97380
gtcctgaaga tgaaaatcct tgcttctcat cttgcctgtt tcagcttatg agttcaacga 97440
accgcttcct caaatactgt aatggatctg ggagttcagc cttgctgggg ccactttgaa 97500
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ttccactctg ttctttaacc tacactggtc ttcctagtta gatggagggt atacattagc 97560
ctgagcagtt aggtggatgg ctggggagca gtggggcaga aggaaaatga gttgggaaga 97620
gtcccagggg tcttggcttg acaatttaag actgagatgg gcaacttaaa tatcctagga 97680
accaagcagg tggcatgaat aaatgaagct ggcagaaaag ggaacagaag caaatgagaa 97740
agcccacacc ctctatagag gattagcagc tactgagctg cagcagattc cagccataca 97800
gatattaaag cccagtttgg ccagatctga tttccaaaaa aaaggcaaaa ttctggatat 97860
ttatatgaaa tctctcccat ttttaatggt ctaattctta tgtattctta tagagataca 97920
atttacacat gtggtaagta gaataatggt ctcccaaaga tgtccatttg gtaagtacat 97980
gtttagcttg atgagaaact actaaactat ttctaaagtc actttcccat aatcctccaa 98040
tgtttaaatg ttggcagcta attcaggaag tttcaaagca aggtataggc ctaacagaac 98100
acacgtgcag gtcagatgtc agatgtgtcc caatgttaag gagagctgca ctggccagga 98160
gaggaagaca cgggtgatgg gcctgtgctg atggagaaag atgtgtctct ctgggagcct 98220
gaatagggga agattctcag aattaggaga aaggcaggtg ggatgcccat gatcactggc 98280
tgtgtctggc aagcagttgt gtccttaaac caaaggaagt taagcgggcc ccacaggaaa 98340
agagttccag ggaccagaga gagaaagagc ttggtttcag acagagattg gaagtgatga 98400
cagaaagggc ctgctcaagg aaaggaagga cctcaggggc cttccgggag ggatggaaag 98460
acaccaaaca tctgattggc aggcgatgac accttacatg agaagtaccc ttggtgattt 98520
ataaagattt tcagaaacat tatctcatga tgcagataaa agccaaactg catgacggcc 98580
tgtactgcac aggggctggc caggcagaag accccatagt gaacaagcag ctgggccagc 98640
tccttgcagg agcaccttcc ctctgccata tgtggtctgt gctcgaggat actccaagcc 98700
agggtatggt ttctgtcttg tagtggaccg agggtggggt tccacaaaca tagagaaaga 98760
aaagctgtag aaggggcatt tcccccctgt ctggctcaaa gggggttctg gaggcaccag 98820
taacccagtg ccttcctagg gcatcctacc accctctcct tgtgcctgat acacatctct 98880
atcttacaga attgtcttta taattgcaat aaaataataa atatgtttct tcagaactct 98940
aggcaatttc tggactcttt gaggccagag ctgaagtctt agttatcttt ataacctccg 99000
tggcactgaa cacaatgcat gagccagata agcccacggt gagtgttttg ctgacttgag 99060
tcacagagcc ctgtaataaa caggctactc aggggtgtca gtcccctcca cacatctggt 99120
tgagacagtt ccatctagtt ctgatcagtt ttattaaata agcttgtgct gtggggctgt 99180
tttttccttt gtgaactggc atttccagaa tgggttacca cgctggtggt ggccctagct 99240
gatgggcttg aaggggaggc attcatcttg tcccatcgcg gctcacttag ctgagacctc 99300
tgcaggagct tacacgcggg ccagcagaat cgcaaatagt tctcccacca cccgaggttt 99360
tatcattgct aagctactgc aagattattg taattgaaat ggttctcagg cttctggggt 99420
tctttctctc tatctctctc tctctctctc tttctctctc tttttttcta gcctgtgact 99480
cacatttttt tctaaggcat tgtatcagtg gcctttgcag gctgctcaag ggatccagac 99540
ttttgcattt attgaacatg tcaggcattc attaagtact gcaccatcag ggagaatttc 99600
cctgctatta agcgtcgact ccagccaaat cagattcacc tgggcgtttc tcaaagaaaa 99660
gcctgtcagc atgataatgg tttgaacagg gctgagctgt atggttgaag tgcgacaggg 99720
cagaagacag gctgtggtga ccccagagtt ggtctgtaac tcacagagga atcacacagt 99780
ggagaggctt cagaaaatca gaaatctgac tgtcacaggt agcggggcaa gctgcctgat 99840
gatgcacttc aggaaggctc agcaggaggg tggggaggat ggaaaaaccc gtaacggagc 99900
agctagaaaa ttaaggcaag gaaaccgtaa gtctgccctc cagactgcca cggttttatg 99960
tttgagactg aggtcactcc cccagggaaa acaagaggaa ctgagaggtg agagtagggg 100020
agaaacaccc ctcactcacc ccttgaataa aaactttggt gtcagtagca tcctgtacta 100080
atgtttattg agcatttatt atgtatcata cttcattcca agaagtgaag gtatattctt 100140
taatttttac aactcttcta tgaagtacgc attaatatta tctatgtttt acaggtgagg 100200
aaactgaggc cgggaagtta aatggcttgc cctaggtcac ccagctagta aggtggagga 100260
gctaggattc agggccatgc catgcctcca gataccacaa ccttagtggc tttactatgt 100320
ggttttccat tccattgggc tgacacccaa gaagccacag gtactcactg tggcccatgc 100380
ccagctttct gtcaaacatt cctcaagttc aagggcaatg ccttttcccc tcctcccctg 100440
cccacaaaac gtgtcctcca aatggcccag ctgagaaagc agggaaattt ctagaaatat 100500
ttgcataaac tttgggtgaa agtggaaacc ccaccaacac ccatcctcca tcctccccac 100560
tctgctgcct atgtctgcca ggctgaggat cctcaccctg gtgcgggacg gccatgtggc 100620
cattcagagt tcttcctacc cccatccaac tttgcagtgc tgttggttgc ttcagagctt 100680
gaggatctct ggacagatag gatttgctct caggtttcac attgcttcct tccttcaggc 100740
tggacaatta gaaatgggta ggagggtagt aaaatggatg cgatgaacta ttccttccag 100800
gcaggcagcc caagctgttg gccctcttct ttccatttgg acttggagca agtccttcag 100860
tggggttagt tactaggagc catgtcagct gaccctcact cctttaagac tttcccacct 100920
tcatgctcac ctgtacataa gttctttgtc actttctgat gaagtctgag cctccaaatt 100980
ccaaagctaa agtgtagtta gcccctacca tttcctgttt agacagggca agatggcacc 101040
cagcagagga gagagaggac atatatgtgg acaatgccct tatcccctgg ggggcttgca 101100
31
tataaaaaat aatattataa aaaattaact agaagcatag atgagcacac acatagt
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
taacaagggc gaaagccctc cctctttctc aaagtcatgc tgaggctgga cctcaataac 101160
tgagaaagaa gagtgacggg gagggtgaag aggttgggga gaggaagctt gaactaaaat 101220
tctgaagtac atgagatatt gtatgctggt gcagtcattc tggagaacaa tcaatgatgc 101280
ttgtcaaaat atgtaaatat atacacacca atcaagcata cccccatctc tggtatatgc 101340
ccggagaaat tcacatgtaa gtcatgagta aggatgttca caacatgttg tttgtgaaag 101400
gagagagtta gagaatctgt ctgtccatca ctacgggaat gggtatgcac aaggcagggc 101460
agtactttgc atcaattaga aacaatcatg gtaatttgat acatcataaa agcagctgag 101520
tgtaaaggaa gacaaatgtg aggactatat cgtgttgtca ttatgtaaat gaaaagacac 101580
aaatttatct tgcaaggatt tactcacatt caaatccaca attaaaacac attggctatg 101640
atggcagtca tggaagagaa tgagagtgaa gaatgttcat aaaagggata attaacaatg 101700
caattatttg cacaatgctg tatcaacagc atcaatgatg tggtaccatg atatggtcat 101760
ggggagggta atgaattcag ttctctgcac ctaaggttga aaagaaaata aaatgtcaag 101820
agtttcattg gagttcattt taattattca tgtaggcaat attgtctaag cataagattt 101880
atgaagaaca aataacatag catggctggg cgtggtgatc caagcacttt gggaggccaa 101940
ggcgggtgga tcacttgagt ccaggagttc gagaccagcc tggccaacat ggtgaaacca 102000
tctctactaa aattacaaaa attatctggg cgtgatggcg ggcacctgta atcccagcta 102060
cttgggaggc tgagacagga gaatcgcttg aacccaggaa gcagaggttg cagtgagcca 102120
agattgcacc actgcactgc agcctgggtg acagagcaag actctgtctc agaagaaaaa 102180
aaaagaaaag aaaagaaaga aaggcaagaa agtaagaaaa gaaaaggaat aacatagtcc 102240
tggactatga atcagaggct atattttctt acttacacct ttgcaccctt gggttctcga 102300
agcctcagtc ctgacacaca ttaactagaa aatgtttatt taattctaac ataacatcct 102360
gagttcctgt gtaagtaatt tgcaaaagcc aaagaatgga ttaggaactc tctgaatgaa 102420
atttctctgc ccagtgcagg aacctgcagg gttaattggt gcctgtgaat acagtatagc 102480
cacagtcatt ttccaaggtt acaaaacacc ccctcgtcag ttcagttgtt tttctttggc 102540
cttcaaagac ctaagtgtta cagttaccat tctgtgcaat agatcactaa agtttattct 102600
tcatgtccta ctgaaatttt ttacctttta ataagtatgt gaaatgatgg atttattagt 102660
tagtttgatt taatcatttc acaatgtaaa catatgttac aatatcacac tgtaccccat 102720
aaatatatta atatatacag ttattatttg tcaatttatg aaaagatcca ggagttaatg 102780
tactgaggct tgctgcagct catggctgat tttatttatt tatttttttt tattatactt 102840
taagttttag ggtacatgtg cacattgtgc aggttagtta catacgtata catgtgccat 102900
gctggtgcgc cgcacccact aactcgtcat ctagcattag gtatatctcc caatgctatc 102960
cctccccgct ccccccactc caccacagtc cccagagtgt gatattcccc ttcctgtgtc 103020
catgtgatct cattgttcaa ttcccaccta tgagtgagaa tatgcggtgt ttggtgtttt 103080
gttcttgtga tagtttactg agaacgatga tttccaattt catccatgtc cctacaaagg 103140
acatgaactc atcatttttt atggctgcat agtattccat ggtgtatatg tgccacattt 103200
tcttaatcca gtctatcact gttggacatt tgggttggtt ccaagtcttt gctattgtga 103260
ataatgccgc aataaacata cgtgtgcatg tgtctttata gcagcatgat ttataatcct 103320
ttgggtatat accaagtaat gggatggctg agtcaaatgg tatttccagt tctagatccc 103380
tgaggaatcg tcacaccgac ttccacaatg gttgaactag tttacagtcc caccaacagt 103440
gtaaaagtgt tcctatttct ccacatcctc tccagcacct gttgtttcct cactttttaa 103500
tgattgccat tctaactggt gtgagttggt atctcattgt ggttttgatt tgcatttctc 103560
tgatggccag tgatggtgag cattttttca tgtgtttttt ggctgcataa atgtcttctt 103620
ttgagaagtg tctgttcatg tcctttgccc actttttgat ggggttgttt gtttttttct 103680
tgtaaatttg ttggagttca ttgtagattc tggatattag ccctttgtca gatgagtagg 103740
ttgcgaaaat tttctctcat tttgtaggtt gcctgttcac tctgatggta gtttcttttg 103800
ctgtgcagaa gctctttagt ttaattagat cccatttgtc aattttggct tttgttgcca 103860
ttgcttttgg tgttttagac atgaagtcct tgcccatgcc tatgtcctga atggtaatgc 103920
ctaggttttc ttctagggtt tttatggttt taggtctaat gtgtaagtgt ttaatccatc 103980
ttgaattggt ttttgtataa ggtgtaagga agggatccag tttcagcttt ctacatatgg 104040
ctagccagtt ttcccagcac catttattaa atagggaatc ctttccccat tgcttgtttt 104100
tctcaggttt gtcaaagatc agatagttgt agatatgcgg cattatttct gagggctctg 104160
ttctgttcca ttgatctata tctctgtttt ggtaccagta ccatgctgtt ttggttactg 104220
tagccttgta gcatagtttg aagtcaggta ttgtgatgcc tccagctttg ttcttttgga 104280
ttaggattga cttggcgatg cgggctcttt tttggttcca tatgaacttt caagtagttt 104340
tttccaattc tgtgaagaaa gtcattggta gcttgatggg gatggcattg aatctgtaaa 104400
ttaccttggg cagtatggcc tttttcacga tattgattct tcctacccat gagcatggga 104460
atgttcttcc atttgtttgt atcctctttt attttgttga gcagtggttt gtagttctcc 104520
cttgaagagg tccttcacat ccccttgtaa gttggattcc taggtatttt attctctttg 104580
aagcaattgt gaatgggagt tcactcatga tttggctctc tgtttgtctg ttgttggtgt 104640
ataagaatgc ttgtgatttt tgtacattga ttttgtatcc tgagactttg ctgaagttgc 104700
32
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ttatcagctt aaggagattt tgggctgaga caatggggtt ttctagatat acaatcacgt 104760
cgtctgcaaa cagggacaat ttgacttcct cttttcctaa ttgaataccc tttatttcct 104820
tctcctgcct aattgccctg gccagaactt ccaacactat gttgaatagg agtggtgaga 104880
gagggcatcc ctgtcttgtg ccagttttca aagggaatgc ttccagtttt tgcccattca 104940
gtatgatatt tgctgtgggt ttgtcataga tagctcttat tattttgaaa tatgtcccat 105000
caatacctaa tttattgaga gtttttagca tgaagggttg ttgaattttg tcaaaggcct 105060
cttctgcatc tgttgagata atcatgtggt ttttgtcttt ggctctgttt atatgctgga 105120
ttacatttat tgatttgcgt atattgaacc agccttgcat cccagggatg aagcccactt 105180
gatcatggtg gataagcttt ttgatgtgct gttgcattcg ttttgccagt attttattga 105240
ggatttttgc atcaatgttc atcaaggata ttggtctaaa attctctttt ttggttgtgt 105300
ctctgagctc atggctgatt ttatttcttc tagtcttggg aagtttttta ttgaaatatt 105360
ttttatgtca taacaatgaa attgcattaa ttcacaacat ggagggacaa gctcttttag 105420
ggagacttct atgtatataa ttattttaaa caaatgttat ttgcttaata tgtcatggat 105480
ttttataagc cataatgtat ttgagcatat cagtttatag tatatgatat ggtgtgttat 105540
tcaataaata tttaaatgtg aaggaactag taattagaaa cacctatgac atgttcctag 105600
aactgcctca cacagcaatt taatattttc tctttttgct gcaacaaatt aagtgtatca 105660
gttcacactg agcaataggg tcactattat atcagtctgt gcatgtttct gaaaaaaatt 105720
attactcaaa ttctaaagca cgatttataa ataacatgtc cattttatca aactcagtgt 105780
aacatattcc aggtgttctt tcaatgatct tatacaaaag atctggcaag ttttcttaga 105840
ggaaagtttt ccggagtttt tactcagtaa gaatcaaatg ctttatgtct ttggtactgt 105900
gattccagaa ttggaggaat ccaaagggat gaagttttat tctgatatat tattccactt 105960
catcataaca acctgatttc agccttctgt ttctcctggt tgcccacaga atcactgcca 106020
ccgtccttaa aaacaagcat tctggccctt tgcaagagca ttataaatca tggtttcatt 106080
ttagggaaaa ggttattgat ttgctcattt tcccctgcct ccctctggtg cagacttcag 106140
tgaaaaactg aagcttcctt tatgaagtat atcatagcca atcaaaagtg tggcctgtgc 106200
ttacttgatt tataatgaaa ggtgacacga gtgagggatt tcacactgct catcctacag 106260
tgactgtaat caagagataa aagctagagc aactacaaat gctggccaga cttcatcaga 106320
attatttact gcgtttgcta agacagggac attcgatcag tttagatgga agaactaaca 106380
cagttaaagg cgatgcattt gtgtttcatt ttgcttttaa agccagctaa cattccactt 106440
ttcctatcgt aattaacttg gccttttaag attgtaccca attgtgattt gctttaagtg 106500
attttttttt tcacaaaagg atcagaaaaa agacaaaggg agaattatcc atggagaaaa 106560
gggcaaaaaa aaaaaaaaaa ggaagtaaag tagtgatgag acatttaacg atatttttga 106620
aaggcctaaa gtcaaggaat aagtgaatat cattagtaaa atgcagaaat gccatgaaat 106680
gtcttggcag caaggtaaat gaggccccgt tagattttct ttctggttat gttatggatt 106740
tttctaaggc atgatataca ttggttggta aagggaaagc cagcaaagct ttctttatca 106800
gtggtgaaag aaaacatgtc tgaagctggg acatttgcag tatgatttta agtatattta 106860
tagagagatt gaaattacaa ataaaaataa taaaaatcat atcaatgtaa acatttacat 106920
aggaagagta ctgcagtgtg attaaatgtt tgtagtgtgg ccgaaactca gaacacagaa 106980
tgctgtcact gaggtgtatt gtgaatgctt atatggaaaa tcccaggcta tcgatttcac 107040
atggcctttg cttagtgcaa taaaaaaagg gattaagatt ttttttcaag cataataaaa 107100
catctgtgga ctcattttca gatctagaga aaaacatcat tattttaaga atccatattt 107160
tttttccttt ccatttagct cacacttact tagtacattt cattaattgc gtctcctaca 107220
gctgagcatg actctttttg catctaaatt gattaaaata gagaacagtt ggtgatatga 107280
gttgtcttcc ccccaccggt tctatatgaa attgaataat ttatgtattg tagaagacag 107340
agtgatctta atatagaaat tgtatttgca gagctcagtg gaggttccca tgttttctta 107400
caaagaaact gatcttttgc cttagctgag ccatttcttc tctaagataa aatgaggtaa 107460
cttttttgtt ggcacatgct ccacacacca tgtcagcagc atacccaaag acttctaaga 107520
caactcttta gttatcaaac aaaactggct ctgtggcaca atggatagca cgtaaaagtt 107580
agttcatagc ttcagaatac attttacctt ttatagtcaa tattgaaaat caaagtaaca 107640
tgtccatagt cctaaaaatc aaatacacta ggcctgggca tggtggctca cacctgtaat 107700
cccagcactc tgggaggaag gcaggtggat cacttcagga gttgaaggcc agcctggtca 107760
gcatggtgaa accccgtctc tactaaaaat acagaattat ccaggcatgg tggcaggtgc 107820
ctataatccc agctactcgg gaagcttagg tgggagcatc acttgaaccc agggggcaga 107880
ggttgcaatg atccaagatc gcatcactgc actccagcct gggcaacaga atgagaccct 107940
gtatcaaaaa aataaaataa aataaagaga gagagagaga aatagagagg agggagatgc 108000
caggttcttt ttaacaacca gctcttatcg gaacaaataa agtgagaact cactcatttt 108060
ctaccactgc agggagagca ttaatctatt catgatagat ctacctccat gacccaaata 108120
tttcccatta ggccccacct ccgacactgg gggtcaaatt tcaacatgaa accaattgat 108180
atggtttgac tctgtgtccc cacccatatc ttgcctcaaa ttgtaatccc cataatcccc 108240
acatgtcaaa ggcgggaaca ggtggaggta attgcatcct gggggtagtt tcccccatgc 108300
33
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
tgttctcatg acagtgagtg agttctcatg agatctgatg gttttataag catcgggcat 108360
ttgccctgct tgcactcact ccgtcctgat gccctgtgaa gaaggtgcct gcttctcctt 108420
tgccttcaat catgattgta agcttcctga ggctttccca gcaatgtgga actgtgagtc 108480
agtcaattaa acctctttcc tttataaatt ccccagtcgt gggtatttct tcatagcagt 108540
gtgagaatgg aaaaacacac caatctatac caatcagtaa tcagaattta cattattctg 108600
acttctactg aaccaagtag tatattgagt atgattactt tctcatacaa cttttgtctt 108660
tcctggagtt aacaattata ttgtgttttt atttgcctgg ttttctatgt gtgtttcaca 108720
aattcttccc atatctatag tctcataatg taggttttca acataatgaa atgtattggg 108780
tgaactatct taccaccatt tttttactga agacatttct ccgagagttc ttggtccttc 108840
cgtttcttag gtctgctgca tatctgtcat cctggaactt ctctccactg ctcccctgag 108900
ttgatctatt tctggaaccc ctggactttc tgacttatac cttctttttg gtgaagtata 108960
atctttggca atttctatag aaaagttaaa cagaagtagg ttgctgttgt tattgtgttt 109020
tggtccttgc ataactgaca gtgtctttat cataccacat actatattga tagtatggct 109080
ggacatagaa ttctaggttg gaaatcattc tacctcagaa tgtttgaaga tattgcccac 109140
tgtcatctag catccagcat tgttgataaa tctgatgcct ttatgattcc cttggcttta 109200
tatatgacct gtattttctg tgagttctta tctactcttt atctgtggta tcatgatcat 109260
aatattcaaa tgtgtctgtt actttctctg cactttgctg agatttattg agccatttta 109320
atttgaaggc ttatggccct agcccagagg actaacacac acacacacac acacacatac 109380
atacatacac acatactcac acacacacac acacacacac acatatatat acctcaccac 109440
catttttttt gttgttgttc tcactttctg gaatgtccgt ctaatgggtg ttgaactacc 109500
tggatccatt ttctaatatt ttgatctcct cattcctatt atccatccct ttgtcttttt 109560
attctccttt ctaggagctt ctttatatct tccaaccatt ctattgagtt tttatttctg 109620
ttattactat atttttattt ctaaggaggg gtgtgtgtgt atgtgtgtgt gtgtgtatgt 109680
gtgtatgttc tttgcccttt ttaatccaac ctttcattgt ttcatggctg tatcatgttc 109740
tcctacatct ctaagggatt gactatgagg ttttgggatg ggagtgatcc gcgttttgtt 109800
ccctgtagtg tctgtccttc tgagtttctt tttttctgtt gttttggtct ctctttcatg 109860
ttggacactc tcttagtacc tgaaagtctt cagttatcta ttaatattta agagcgaagc 109920
acggaaaatc tgactgaaaa ctgaataagg ggagcttgtc aactagggtt gagggtgagg 109980
gtgtgtcaca gtcaggcgtt ctagcagcag gcagcctttt aatccaaagg tactctaatt 110040
gtttgaattt gtctcttttt tagaagacta ttttctgtga aaacgaacaa ttcattttcc 110200
atattggagg agtaagcctg actcctgggg ttctgagaag gatgtggagg ggaatgccac 110160
actattcaca ctggtgaaaa ctttgactta aatcctctac ttttagtctc acacttaact 110220
cattttcttt ctgattgatt tacccaagtc cagactattg caacatgagt gaagggtagt 110280
cctctgattg cttggctggg gagaagtctg aagttctgcc cactgcattc acaggttttc 110340
gatgaatttc attattttca gcctgtgtct cactcctatc ctcctaagta tctggtgcct 110400
tcaacatctg acccttttgg gatcctaggg aataaatagt cttccccccc acttctgtat 110460
gtcctgacac cacagtttcc ttggctccac taagccagtt attcttcctc tgtctgcttg 110520
ctgtcttcca aaatcttgtc gagatctcat ccctcattat ctactctcca gttttctttg 110580
ttcttgttct ttattccttt acccttacat agtgatgttt gaaatgacag aatgatattt 110640
agtctgccat gtccattgag cagtcaagaa taagttctcc atgtggaatc aaaggagata 110700
gcagagcaaa taaatgctgt acttgtgctc tattgctgca taacaaatca ctccaaaact 110760
ccgtggctta aaacaataac catttattat ttcatggttt ctctgggtca ggaatttaga 110820
cagtgtgcag aggagatggc ttgtctctgc cgcatgatac ctggggtctt tgctggcagt 110880
attaaaggct gggggcctag tattatgtga agacttatct aaggctgaag gatccactta 110940
caagggggct tactcacttg gctaccaagt tggtcctggc aagttggttc atctccacat 111000
gagcctctcc ccaaatgggc ctctccaccc tgcttgagtg tcctcactat taaaggttag 111060
caccccctaa ccccaagcta gtgatcaaag agtaactaag gcaaaatctg caatgccttt 111120
cctgacctag cctgggaagt cacacactgt cacttctaca gcattctgtt gccacccatg 111180
ctagccacaa ttcattaatg tatgaatccc aggtgatgtg gatcgctagg ggcatcttgg 111240
aggctagctg tcacaaatgc ccaactccca ggagggctat ctgcttccct tctttgcaac 111300
atgattttgt gttggaaaga agtcagaaaa aatattttcc tccctgtctt aagctaagat 111360
gttggtgttt ttgttccttt tcctttctta tgctttttat tctgcaccat tcatcaaata 111420
ttgtgccctt tttctttgaa atcttgaaag atgaagaatg tgaaagcaaa agagaagaat 111480
ctaatgccta ctatttgttt tggtggcacc cctacagcat aaaattgtgg cacgtgtgag 111540
ggagaacata gtgttggttc agtgccagcc aagggagact gcattcccac acccctaccc 111600
caaatgtggc aagtaataat tacacagcag ttgtgtgata acttctgctt aaagcagatg 111660
tattttgaca catcacctgt cccctgatat cagtgtagct gtgtaaacag tagtgtgtac 111720
attaccaaca ttttacccca agcatcccct tagggtctcc gattttctgc ctcagggctc 111780
tctccaaaga cctctcatcc ccacacagac tcagcctcga agtgcagggg aagtaactaa 111840
ctaggaatca accctcaact aaaagacaac agaaggcagt ggataattgt cctctggagg 111900
34
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
aaccatctgt gacacattct acatggttcc ttagagggtc cccaagtggg attgagtccc 111960
tcttgttcct agcagtaagc agttctgtta tacactcttt atatgctttt cctcctttcc 112020
tctctcagtc tctctgatcc ttccacctgt gctctgggtt cacctctcaa gtaaaccacc 112080
tgcacacaag tccctgtctc aggctccatt tggggaaacc aaactaagac accacctcaa 112140
aacacagaat ttcacgaggc tccaaatgcc tcagaaaagt aatgtaaata taaaatgacc 112200
aattttttga agtatattaa tcctagttat gacactacag gcagatgcca gataaatatt 112260
tatcattgat tagatccact attattaaaa atacaaactc ccagatccca ccccatttcc 112320
actttttttt gagatggagt ctcactctgt cacccaggtt gcagtggcag tggcgtgatc 112380
tcagctcact gcaacctcca cctcccaggt tcaagcgatt ctcctgcctc agcctcccga 112440
gtagctggga ctacaagcgc acgccaccac acccagctaa tttttgtatt tttagtagag 112500
acggggtttc accatgttgt ccaggatggt ctcaatctct tttttttttt ttttttccca 112560
agccaaactg tatccagctt tattaaagat actttccata aacaatcatg gtatttcagg 112620
caggacatgg gcagacaatc gttaacagta tacaacaact ttcaaactcc cttcttcaat 112680
ggactaccaa aaatcagaaa gccactataa aacccaatga agtcttcatc tgatgctctg 112740
aacagggaaa gtttagagtg agggttgaca tttcacattt agcatgttgt ttaacaactt 112800
ttcacaagcc gaccctgact ttcaggaagt taaatgaaaa tggcagaatt tatctgaaga 112860
tccataatct agaaacagaa ccactgctct tttgacaggt gccatctcag tggcatcact 112920
ggaaagtcca gattgcctaa cacactggta accaatgact aggggtcagg tcccaacaga 112980
tgtctgggct taagggagtt aagtctatgc tgaaagaggg aaagggagac gaggacataa 113040
aaacaaattt gtttttctat accacaaggc ttttgtgcca aggtggccat gtgtgtcaaa 113100
gtcagggaat ccctcctcct ggaagccaag aggaagtctc tcaaaactag aagggaaagg 113160
tgttttctcc acatcaatcc agctttggag acattctatt agcgacatat gccccttccc 113220
ccaaaaacaa caatgaagtg ttctatgtgc taacaacata gctttaaaaa aaataaaata 113280
aaataaaaca aaattctgca tttttataaa acttgataaa aaatagtatt tcaaactgta 113340
cagtcaccag aagtacacag ttatcaaaaa tgcacacact tcacttggca tctccagcac 113400
cttcagcttt ctgcgcctgg tctgttttgg catctccatt tcctgcaggg ttatttccct 113460
ccttgccagc atcagctttt ccctttttcc ctttgggtac cttctctccc ttctttgcag 113520
gggcctttta ggcgtgggct ctggctttgg aggagcaggt ttagcagaca acctcgcgga 113580
tcttctctgt ggttcgtcct tcaccttggc tttatctccc ttagcatccc cttcagcctt 113640
tctcttgggc atggtggcgg cagcgacggc agcgggacat aggtgctgga cgcgggacgc 113700
agcggcgcgc gggctttggt cggaccgggg gtcgttctcg cctcttcttc acactgctcc 113760
ggtctcaatc tcttgacctc atgatctgcc cacctcggcc tcccaaagtg ctgtgattac 113820
aggcatgaac caatgcgccc agcccagttt gcacttttaa taaggaaccc tggtgattct 113880
tatgtaggtg gtttaggaac cacactttaa aaaacgctgg cctcatggct tcaagtgaga 113940
catgttttgt tttattggta cagaaacaaa atgctgcgag gtcacacaga tagtgggtac 114000
agggctagag ttaaaatcca ggtgtgtctg acttgaagcc attctctgca ctgaccatct 114060
gcataatctt ggagcatttt gtttcttcat caagaaaacg aagatgatgc caggtgtggt 7.14120
agctcacgcc tgtaatctca gcactttagg agcctgaggt gggaggactg cttgaggtca 214180
ggagttcaag actagcctgg acagcataac aagaccccat atctacaaaa aatttaaaaa 114240
attagtagta gtcccagcta ctcagaaggc tgaggcggga ggatcccttg agcctgggag 114300
tttgaggccg cagtgagcta agatcacacc actgcactcc agcctgtgca acagatcgag 114360
accccatcta aaaaaaaaaa aaaaaaaaaa aaaaaaagac aaaaagaaaa gaaagaaaga 114420
aaacgaagat agtggcatct acttgttagg tcattactag gataaaatga gatgagccat 114480
gtaaagaggt tggtgagggg caaggcacca tagcaagtgc tcagtaaatg gttgctgtgc 114540
acacacagag ctataatata cttgcactac agaaatgcat ggaatcaaaa aaaatgtatg 114600
ttttctaaaa acagagttaa tctgctaggg acttacaaac ctcaagtaaa ggattccaca 114660
agaacgctct cagattagtt ctacagtcaa caaatgcgta ttgaggggat tttatttcct 114720
ggtataactc aaagacttcc tctgagggtt tttcagctga gtgcaggaca gcagctggga 114780
gcacagaggc agtccacatg agcattttct gggcttgcca taatgctttt gactgggaag 114840
agagattttt ttaaaagatg gcttttcctg gatctttttc aatttgtagt aaaatacaca 114900
gtgggattct gaacacatct cattttattt tcctgctgca tacacccttg gatgctcaat 114960
aagtcactgc tgactggtgc caaagcctcc tgcctgctct ctttgattct gcttctgtgc 115020
cgtgccccgc agtctattct caacaagaca accagtgtgg tccttcaaaa atctagatct 115080
cttctgtcat ttcctattct tgaccccctg tgtctcctca tcacacttga aagtaaacct 115140
caagtcctgg ccgggccctc cttgggtgtg cccaatcagg tcctggctta cctcacatcc 115200
tcccactcca actcacatgc agccacactg gccttgctct gctctgaata cgccaaaaac 115260
atttccctct cagggcattt gcacatgtag tcccctctgt ttgaaatgtt ctctcaaaca 115320
tccacaggga ccgccctctc tctgcattca gttctctgtg gcttggagag gccatccctt 115380
cccaccctac ctaaaatacc taaaatagca cacccattct cagtccaagc ccctagctcc 115440
ttacactgat ttataacaca cattcaatat ggcatcttca gttagaatga aactccatgt 115500
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
gggcaggact ttgtctcact tactactctg tccccaggcc tagaaaagaa ccaaacacgg 115560
taagctctca aaaagcagaa tgagatgtca aatgaagtat ttctggtgtg gattggggct 115620
ggccacatgt aggatatctt gcttaattga caaatgattt atagagcccc tgttgtacac 115680
aaaacatggt gccaaggcaa aagatgagta agaccaagtt cctgccttca aagagttttg 115740
gtcttgccca gtgctgtcca gtagaaatat aacatgagcc acaaatgtaa tttaaaattt 115800
ctagtaacca catcaaacag taaaaagaaa tatgtaccat tagttcttat aattacctta 115860
ttttaaccca acaaatatcc aaaatgtcat ttcaacatgt aatcaatata aaaatgagat 115920
agtttacatt ctttctttgg tattatcctt taaatctggt atgtagttta catttacaat 115980
gcatttcaat tcagatgcaa gtttttattg gaaacccttg ttctatattt agagtccata 116040
aagtttacaa aatgtcagtt catattcaag ttgatccaaa catacttaat ggtttttgaa 116100
tcactgaatc aagttaaatt catcaaaatt aataaaagtc ctaattctgt ttctcagtca 116160
cactagccat gtttcaaggg ctcaatacca gtggctagtg gctgccatat tgtactaggc 116220
agtccaggtc taacagaata gatgaggtgt ataaatgtgg ggaagtgaat acattcatga 116280
agagatcata taaagaagag agtcaaagtc tgaatactgc ctttgtacac aacctcttcc 116340
tgacaggaac tgtctctttc tttaggactc aatttatatg tcagcttttc taggaaatct 116400
tccatggtac ccccaaattg ggttagttcc cctcaagtgt aagatcccag agtaccctgt 116460
atgtctacca tcttcagttt gcccttctat gtttcaattg ctctctatat atttatattt 116520
gcatttcccc cgagtttata aacttcatgt ggccaggagc tgtcactgtt aattcctcaa 116580
tgccagattc ccaataaatt ttgtaggaat gaatgaatga atgaatgaaa tggccaagac 116640
ttggaggaag aggaagagga aggactagcc ctgctagaat ggagaaacag gctgtggaag 116700
gggttccagg atggggcaag gaccagggat gaagatagaa gctcagttca tgcaaaagtc 116760
accagtggca aatgattctg aagagtccaa ggagatgtga gctaagaaaa cacgatcaga 116820
cttaataagc aggaagccac ttttatggga gcctacaaga gacaatcaga tcaacctaag 116880
ctcaatactt tttttataag ttcaaaaatg tattgatgtc cttggtttaa gagtttttga 116940
tattttaaat attgtgagta gactggattc taaggccgtg gtccagtttt aatgtctggt 117000
cagctgaggt tcttctagtt actgcagtct ccaccaatga cagctgttaa ctctatatgc 117060
taattgaaat tgggagcgaa ttgcagctgg cttggggcat ttcgtgtgat ttatgggcat 117120
ctgatctaaa gatacttaaa ggatggaatt attggagcca ggaagagaga tgtgattaat 117180
caaagaagac aattcagtaa atggcccact aaaacactgc agtataaact aaaacaggaa 117240
ctctagttcc atctattgag gcaggggagt gtgtgtgtgt gtgtgtgtgt atgtgtgtgt 117300
gtgtgtgtgt gtgtatgtgt gtgtgtgtgt gtgtgtgtgt gttaaagggt ataaacagga 117360
tatagaaggt tttttttcca gcatatagag attatggttg ggggattctg aaggacatat 117420
atttatcttt taagtttgct tacaatgtac ccatcaccat caggaaatgt caagttccta 117480
gccctgtgat ttcatcctga ggactcctgg gagttagggg aatcaccatg tgggaaaaaa 117540
aagacaatac cttttattca aatttttaag ggccagtgtg agttgtcatc tttccatatt 117600
cctgagtatt cattcatttt tatcataagg cctgtgggtc tgcggttacc tttgtcaaaa 117660
ttactggcac taagaaaaca ggatgcaagt taaaagttct cagtatttat ctaaatagaa 117720
aaataagtga ttatcggtaa catataagtg attgtcttga ataattgttt atttgtgcct 117780
tcagagttaa cctagactta tgttcctcct gtccagtgga ctgtcttatt tattgctgaa 117840
gacctaactc tgacaggcag ctagacattt tttcagtagc tgtggaccat taaaacaatt 117900
ggtcagttaa gtcctttcag gaggaaaatc tctgacttgt ttgtaactgt gtaagtaggc 117960
accttctggg tccacacagc acaaatccta aatagcaaca cacacactca cactcacact 118020
cacactcaca cacacacaca cagcagaaga gaaattcttc ttagaaaatc aggttaaaaa 118080
gggaagaggg aaaaattatg cccagcaaag caagacatag tggtgctgct tccctcccca 118140
tagacactaa gactcagtgt ccccacctct gtcaccctcc cctaccctac atatacacat 118200
gcacccatta acatgcatac atgcttgctt tttccacact aacgcccctt cttgtgtcaa 118260
ctctaaagtg aaaagaatct ggggcctaag ttagtggaat actcacagaa atggaagcca 118320
tggatggatg gtatgggtgg aggtacgaag agattcgggt ggcatctttg cttttcctct 118380
gaaactatca aagaaatgat taggaatgat atatgtgctg agagatgtat tgatgacctt 118440
tacagaatgg aaattgaaga accagagatt aaagattaga aactcaaaat cttttagccc 118500
cagagtgtcc tagttcccta attggtaaga caggggccag tgaggcggac acaccttgaa 118560
gaaggactaa ctaaggatca cagtgctgta ttgaccttag caaatgctac agaggagagg 118620
aaataactga gccagaacaa tgattgtgtc ttatgacctg ccgtgtgcaa cagatgggat 118680
gctaaacact tacaggtact tttaattgaa taatgctttc ccgtggccat atgctgtgat 118740
tatttgtatg cttcctgttc atgctttatt catctgtagc tgtcactcac ttaaaaccaa 118800
ctttcccttt ccatatctag ggattgaagt ggatactgtg aagtgtgaat gagaaataga 118860
aaaaacattg tttactgagt acagcttatt ttctcctgca acgtgattat ttattatctg 118920
tgtgttcatt tcagattatg aattattact gctggtaatg cctgttcttt cttccagttg 118980
ctgtgcattg gcacacagga gcgaatatca aatgccaagg ggagggtaat acaacatcgg 119040
ggaactaggg taattctaac aatctattgg gaagaaaatc tgtgcacttg aattttgaat 119100
36
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ttaagtcaaa gtttttaaca attgtggacc aaaagtagtc tttctttttg tattaaaaca 119160
gaaaacagtt gccccaacca aagcctctac aatgccatat cattagcttc tgagattaaa 119220
tttgacaata taagattaca tgcatgctaa tatttcataa ataaataata aggcatgcaa 119280
aagtcagctc tcagccatcc cttcctcacc tggcaaggta tttgatgaca gtgattatta 119340
ggtgactgaa agagaataag caaggccgta atgaaggaaa gaaattaatt gcccaaaacc 119400
tggagagttt ttggttcaga gctgctaatg atcaagttat atgcatgtcc tatttgccat 119460
taaaatgctg caaaactgga aatattctta atggaagaac acactattgt gtactttggc 119520
ctgccacgag catcatctag aacccctcaa atgcatagca cacatgttca taggcagcta 119580
tgttagtttg ctagggctgc cataacaaag tgccacagac tgggtggctt caactgcaga 119640
aactcatttt ctcacaaata tggaagctag aaatctgaga tcaaggggtt ggcaggtttg 119700
gtttcttctg aggcctcttt ccctggcttt tagatggttg ccttcttgct gtgttctcac 119760
atggtcattt ctctgttttc tgtgtcctaa tctgctcttc ttataaggac accagtcgta 119820
ttgaattcag gcccacctta atgaacccat tttagcttaa ttgcctcttt aaaagccctg 119880
tatccaatat agatctgagg tactgggggt tagggcttca atatataaat ttggagggga 119940
ctaagttcag cccataacag cagccatctt tctagcccag tgccctgcac atagtaggtg 120000
ctcagtgatt tttttttttt tttgagacgg ccttgctctg tcacccaggc tggagtgcag 120060
tggtgcgatc ttggctcact gcaacctcca cctcctgagt ccaagtcatc ctcccacctc 120120
agcctccaaa gtagctggga ttacaagcat gcaccaccac gcccagctaa tttttgtatt 120180
tttagtagag acaggatttc accatgttgg ccaagctggt ctggaactcc tgacctcaag 120240
tgatccacct gccttggtct cccaaagtgc tggaattaca catgtgagcc accacgcctg 120300
gcctcagtga acaatttttg aatgactgaa tgaaaagaat tagccagaag ttcctgtgtt 120360
gagagaaaag ggtgggtatt attaggagga gtgtggtgat gctgtggttg caactggacc 120420
attcagagaa aatagcaatc agttttttaa aaatatttat atccttaata aacctggaag 120480
aaaagtaaaa ttctagatcc cctataaaga gacaaggaga aaagacttgg atagtctctc 120540
tttgagggac acagttggaa gtatacacac agaagttctc caagagctgc ttctatcata 120600
tcttgtgatt taaagctgat tgcaagcaat tagcagcact tctgagttta tgaaaatgtt 120660
aagtgggtgg ttctgtagta gattttgttt actctagagc ccattgttta ttttaatgca 120720
ttcaaacaag aaatgcctga tgctagatgt aaaatgttta tcaattttac atctccccat 120780
agtcacaaag tttaaaaaaa gaccatgtga agtaaattgg aaagatgcat gatgcagtcc 120840
cattatccat taaatattca acattggctt tggaaaatat tgtagcactg taatttaaag 120900
ccaatttggg tagcatttta ataagatttg aggcttttga ataattgtgt taataccagc 120960
agctgctttt atttggttag taagatgact aattctataa ttaatatatt taacaaaaat 121020
atactgaaaa tatattcatc ttgtcattaa gatgtccatt tgtacaggtt tctctttaac 121080
atgaaaagat agtgtgagga tatcattgcc tccctctgct atgatgaaaa tgatggcagg 121140
taggtgtgtg ttgctgatgc gcatggctgt gtttttgtgc ggcgtcagtg gcagcactga 121200
gcgagaccca atcgaagact ctgcagagcc atggggtata taaatcttca gagtctctga 121260
ctcagtcaac catacagcag ctcaattttg tgccacaatc ctggtaatga gatttttcca 121320
tggtattggc ttaacattac ccctcaatgg gaacgtttta ttctgaacgt ttgctttgtg 121380
gaatgatctt tcagttctct ctcaaagtca ctcctccaca acatagaaat caggtgtgaa 121440
attcaaataa ttgaagacct taattcttca ttgtgtaccc aagtttcgag ggttactgcc 121500
ttggtaaaga aatgttggca ggtgctccat ctttgcctca ctctcttctc catgtcattc 121560
tggctgttct tttattgcgc gtcctgcttt ttttaaaata tgatgaagaa aagttcaaac 121620
ctaactggta aattaaataa aatacctaac attgctatta tttaacttga tttttgtttc 121680
ctacatgtgt tttaaaccaa aaaaatactt tatattttta aaataggaca ttagggctct 121740
cagcaatgtc ctgactgaaa tcttgcctct ctcctttggc acactgtggt ttattgttgt 121800
tgctgtcatt tcacgttgtt gttgctacct catttgcact aaataagcgt tctgtggaaa 121860
cagcatctga gccactgctt tccacccgtg ttctagatgc atccaatagg ttttactaga 121920
ggaaaggtgc tatgtgtcca cttggaaaaa ctgctagaga cactcttcct ctttgagtca 121980
taatggattg gggttcaccc ctgccccact gacagcagcc tatctgcact tgattgaggt 122040
tgtctgagaa gggtctccgc tcctgggtca gcattagaag cctggagtga aggaggaaca 122100
ggctacactg gctatgctgg ctcagtacta agcagaaaac gatatcatca agagcaaaac 122160
ccaattattt gtagccagat gggcacagat taatttctct tgaaatctca atggacgttt 122220
ttttttcttg tgattctctt tttataaaag agaagattaa aagggcttct tgcccttgcc 122280
tacaagaagg gcagccatca gctgtgtatt atgagagctc ttaagttact tacgatctga 122340
aaactaccag gtagacccca cacctccatt cctggggtcc tattgtttcc ccgacccact 122400
gcagttacca cagcatgtgc atcaggagca tttcccacat ctagggaaat gtagtagcat 122460
gttgattcag aactgattcc ctttaaatgc ttcctcttat gtcatgcatt attattcaag 122520
tcactcacta ttctgagaaa cattgtaggg ttccccacac ataaaacaaa gcatcacggc 122580
agccatgttt tcagactttg gcatgcaata gagtcacctg cagggctcat caaagcacag 122640
gttgtgtcca ggcatggtgg ctcatgcctg taatcccagc gctttgggag attgaggcag 122700
37
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
gaggatcgct tgagtccagg agttcaagac cagtctgggc aatatgacaa gaccctgtct 122760
ctacagaaaa aataataata ataataatta gctgggggtg gcagctcata cctgtcatcc 122820
tagctactca ggaggctgag gtggaaagat gcttgagccc aggagttaga ggcttcggtg 122880
aactatgatt gtgtcattgc cctccagcct ggacaacagt gtgagaccct gtctcaaaaa 122940
acaaacaaaa tattaaacac acacacacag aggttgccag gtctcacccc accatttctg 123000
attccagtgg tctggatgga accagggaat ttgcatctct aacacaaccc caggtgatac 123060
tcactctgct gatataagcg ccacacttta agaaccagtg gtatccagtg tcagagctat 123120
ccatgcacct attctttatt ttattgtttg aagttcagga gaccagggtt ttgagcttgc 123180
taacaaggtc tctttattcc agggactgac atcacttatg tacatgacca cttaatcatt 123240
aatttaatca ttaaactagg attgttagca tgaaaacaac agtacataac actgacatag 123300
aatttgagtt tacaaaacac ttttactcca ttacctcatt caagcctgca gacaagccca 123360
aagattattg ttattatttc tattctgcag atgacaaaac tgaggttcgg aattaggagc 123420
attaaccaga ctaattccag agctttttga tcttcaaact agagtatgca aactcactgt 123480
ccaaggagta tatgcagtga tgcttttttt gttgttgttt ttgagacaga atctcacttt 123540
gtctcccaga ctggagtaca atggtgcgat cttggctcac tgcaacctct gcctcctgag 123600
ttcaagtgat tcttctgcct cagcctccca agtagctggg actacaggct cccaccacca 123660
tgcccagcta attttttgta tttttagtag agatggggtt ttgccatgtt ggccaggctg 123720
gtctcaaact cctgacctca ggtgatccta cccgccttgg cctcccaaag tgctgggatt 123780
acaggcgtga accaccgctc ccagccgcag tgatacattt aagaaagtca attttaagaa 123840
actccacatc cataagcact ctttccttaa actgatctcc ctgagaacac aaccttcttc 123900
taatctatga aagaaagata taattttcac tagcatcaag ttagtaacct ctgggctctg 123960
aacaaagagg tgattcaaaa tattggtata gctggtgaga acgtgactca ctctaaatag 124020
gcaagtaata aatccttttg tggtcaggtg gtttcgaata tttttctttt aacaacattg 124080
aaaaagaatc aaataagcta gtagatcgtt aacaattttg atgatggata actaagtgaa 124140
ctcagagttc acagaattga gtgatgtagc cataaaagta aaactccttc catttgtatc 124200
tattatatga gtaagaatta atgacaaact ctgtctcatt ctatcaacaa gtaatattca 124260
tccatggata catgagccaa ttggggaaaa aaccctccac ttcatgttat taaaagataa 124320
atatccaata caactttatt ttttagttta aaaattatca aaaattgtgt aatacatatt 124380
gttatattga tcaattgtgt aataataact gcaataatgg ctcaatcgat agagttttat 124440
cacttaaaga gcttgtggtc acaggactct taccttttaa atttagatac atgtgtttgt 124500
tgcagagaat tataacaggt tgatcaataa aatcagaaaa agcatgattc atgcctaaaa 124560
atgtactgca ttaggttaga attctgtgca gatagtggaa tggaaattca aggtcaaaga 124620
gaaagaagaa acataaaatc tccttttgtg ggttgatttt tgttttttta tgttttttgc 124680
tttttggttt ttttgagacg agatctcact ctgtcgccca ggctggagtg cagtggcatg 124740
atcttggttc actgcaacct cggtctcccg ggctcaagcg aaaatctcct ttggatcaaa 124800
actagtctgt ggggttttca agtggccagt gacaggttca aaattgctat ggtacttaga 124860
ttccattggc tacattttaa agattaatga catggtttta tttaaagatg ctgttatttg 124920
caatatgcca gaaattttac catttaggat gggagggaaa tggtttctca gaattctttg 124980
ggattagatg agcataaagt gtggttccat attactcacc tctaattact caacacatcc 125040
agccgtttct gggacagttc tattcctttc cccaccacct ctcctcccta aggaaaaaaa 1257.00
aaaaaaaaaa aatctcacct ttccttacta cgaagatttt atctttgtga aatcaatgtt 125160
tcttagttat ttaatatcac cgagtcaaaa tcttttagct ttacctacag gcataattta 125220
acttagaaac gattatctcc atgtattatg agaagaaaaa ctagttcaat tttctcccta 125280
tcataagttt ttaaagattc tgctgcagac tattctcact gagtttcaaa aacttgaact 125340
gcttccctca ttagcacttg gcagaaggta gaaatgtaat gcattgtctg ggtgcaggag 125400
ctcacacctg taatccaagc actttcagag gccaaggcag gagcattgct tgaggccagg 125460
agtttgagac cagcctgggc aacataggga gaaaccacct ccctgtctct attattaaat 125520
atatgtgtta ttatatatat atatatgtgt gtgtgtgtat atatatatgt gtgtgtatat 125580
atatatgtgt gtatatatat gtgtgtgtat atatatgtgt gtgtatatat atatatatat 125640
agagagagag agagagatat taaaaattta aaaataaatg taatgcatct acacatccag 125700
ccagccaatg tggcataata acagcattat ttgcatccct acttttccat ctattccttg 125760
agagtgtcgg tgaaagtggt tctgagagcc ccagctggag tgagacagaa ggcaaacatt 125820
actcaaacct ccatgtttag ctttacaatg gtgtcactca cttgatctca ctcttttagc 125880
ccttatgaac tgcctatttt ccatggaaat ttccaggcat ggaaggtaac atctgacata 125940
gatatgccca aacattcaat gtgaaataaa tacacagact gcctctgatg tgttctgaac 126000
cataattcag atttcaaaaa tcttaatgaa tgaagcttct actttcccca agactgtcac 126060
taaatcttct catggcttaa ttaaatacta ataattgtac taaatccaaa atgttaatta 126120
tgtttatctc tgagtgatga aacttggagt tatttacttc ttttattttc tgtattgctt 126180
gccacataga catgtattat tattgtaacc aagaaacaat cacattgcta tctgagggta 126240
gaagaaaatg atcaaaagca tctgagacag gcatattggc tggggtccac cccttttcct 126300
38
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ggtacaaatg agactctaag tgttgattta tatttatttt ttataaatcc tctctccacc 126360
ccacccccaa atggatttta ctttgtctac attttttaat actcactgtc aggaaatact 126420
ctgttaacct aagattttga gcttttccct attattctgt cctatggaaa gaagaacagt 126480
tattcatata atattaatga tcactcttaa tatcattaaa aagctctgtg gatctctcca 126540
gtaatttcta atacatcaag gtagccaaag attgttttgc agataaattt attttcctgc 126600
cagggatttt taaaaccata aagcattatc aaaatcataa agtatatgtt tctagctcac 126660
tcttttgaat ctgtctgttt tcccagactg gaattcacaa acgcattctg gttataatga 126720
tagcacgtgg tttggagaca ggatcagccg gtctggagac catcttccca tgtcacccaa 126780
gcctcccatg cagggaccct gtcttcctcc accccagaat gtccatggtt gtccctttcc 126840
tttgtccctg gttctagagt ccttttctca ctagtccttc aggaatgtat cacatttcct 126900
tcttaactgt ggccctagag tggtcctttg gccccactgt cctagcttgg atctctgtct 126960
gtctagccag gcatcccttt gctctattag attttcccta ccaaatccca gtccgtgggt 127020
tgccatcaag taacccttac atgtcagcac cgtgcctgtg aaagccaagc ccctcacctg 127080
gtgaaatcgt tatcagagtg gtcacgcctg gccaccttat gcgcccccct cacccctcag 127140
gttctgctga gccactgggc cactggagga gggactgcct cacctgagac tctcatctcc 127200
gcagccacct cccaccaaat gcccagttct tcctgcctct tccttcaact tggctcccat 127260
ctcagccctt ctgggggaag agggctcaga ttcttacaat ctccatagaa aggaaataca 127320
gtgtcaggcc agtttgcctg atgccttttt aaaaatcaat aaaggggaca gaatgcaaaa 127380
tatgtgctag gttgttttta tgaaatgcaa ggcaaaaaat gccccattaa acatttcttc 127440
tctgcagccc ttgctctcaa gggcacatgc ttgacccttg ggttcttaag actttcacca 127500
aggccacttt gtgctttgcc tgctctgcct gagtctctgg ccctcagagc tagagggtgg 127560
ggctggcctt ccagtctgta tctgtcccct gggggagtac ctttccaggc tgaatttaag 127620
aatgcagcaa ttgacttaga atgcagactc attgagtgaa tgcgagccct ggaggaaaga 127680
aaaatcactg tcacttctgt ggcccccagg tttgacactt tcagggcaac cccagagtct 127740
gaattgcttg gaacatccac tcacgcatgc aagtgaaagt catcagggaa gaggctttta 127800
atagcgcagc gtctgtgtgt ctcttgtgaa cccagggtgc cctcaagtgt tggctagggt 127860
taatcacaag ccgctgtttc tgagaacacc ctagaatgaa gcaaccctta aacgttttag 127920
ctttgtgact tgtgtccatc actcttaatt ttctgcagaa gctttttcct gcctttgtta 127980
cttctgtaaa ttcagtctat cagagggctt atgtggtact tttaaaaact cactttgatt 128040
tgcctttgtg caaattgagc tctgaaagag agcagtttcc agggaaattc atctcccagc 128100
aagaatcctg cctgctacca aatcatcttt taatctgccc cccgctttaa aatgtcatta 128160
tattttcctt atacagacct tcataaaact tacaaacact gtctagttta catcttcatt 128220
ttgctctcac aaggccaaaa aacttgttaa acagacttaa tgaacgagtt cctcttgatc 128280
ggggttgtaa gtttctaaaa ttgcatttga acttctacct gcctctagct gaatgaaaat 128340
tgcctccaga catctgaagt ctttttcttc ctttcatttt gcttaatcta cctctgtatc 128400
aaagcacatt tccattttgt ttaggccagt tgtcagactc taggcatttg cagattttag 128460
aaacctgcac aggtgttctg atgtgccgta aacttccaga gccactgaat tcagcaaaga 128520
aattccttga tgttcctact tattaatctt attaattaac atatatctta tatattaatc 128580
tttatactta ttgttgtcac tggggctaaa tagtctctat agctcgtggt ggggggctgt 128640
agtgtatgtg tgtgtgtgtg tgtgtgtgtg tagtgtgtgt gtggtatctg tatggtgtgt 128700
agtgtgtgtg tggtatctgt atggtgtgtg tactacatgg ttgtatgtgt tggggtgtgg 128760
ggggtgcgtg tagtatgtgt ggggtagtgt ggtatggttt gtgtgtgggt aatgtccagt 128820
atatgggtgt gtgtagtgtg ggggatagcg gtgtgtatgg gtgtgtggga gtgtcctgtg 128880
tgtgagggtg gtgtgtggtg tggtgtatga gcgtgtggtg tatgagtctg tggtttgtgt 128940
atgtgtgggg ggacatggat atgtgtggta tggggtgtct atagtgtcag cagatggtag 129000
tgtgtgtggg tgtgggtatg taggtgtagt gtgcatggaa gtgtggcgtg tgtgtggtgt 129060
ggtgtgtggt atgtataggt gtgtaggtag gtgtatgtgt gtgtatgagt gtgtagtgtg 129120
ttgggtgtgg acgcatgtag tgtgtggtat atgggtgtat gtagtataga tggtgtgtcc 129180
tatggtgtgt aatatatggg tatatgtagt atggatggtg tgtgtggtag gtgtatgtgg 129240
tgtgtgtgtg cagtgtgtgg ggtttgcaac atgtgtggtg tnnnnnnnnn nnnnnnnnnn 129300
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 129360
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 129420
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 129480
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 129540
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 129600
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 129660
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 129720
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 129780
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 129840
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 129900
39
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 129960
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130020
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130080
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130140
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130200
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130260
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130320
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130380
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130440
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130500
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130560
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130620
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130680
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130740
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130800
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130860
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130920
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 130980
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131040
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131100
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131160
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131220
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131280
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131340
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 231400
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131460
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131520
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131580
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131640
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131700
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131760
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131820
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131880
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 131940
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 132000
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 132060
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnntgtt gttggtgtat aggaatgctt 232120
gtgatctttg cacaatgatt ttgtattctg aggctttgct gaagtcattt atcagctaaa 132180
ggagtttttg ggctgagacg atagggtttt ctaaatatac aaacatgtca tctgcaaaca 132240
gagacaattt gactttctct attcctattc aaacatgctt tatttatttc tcttgcccga 132300
ttgttctggc cagaacttcc aatactatgt tgaataggag cggtgagaaa gggcatcctt 132360
gtcttgtgcc agttttcaaa gggaacgctt ctagtttttt gcccattcag tatgacactg 132420
gctatgggtt tgtcacaaat agatttacta ttttgaggta tgttccacca atacctagtt 132480
tattgagagt ttttagcatg aaggagtgtc gaattttatc aaaggccttt tctgcaacta 132540
ttgagataat catgtggttt ttgtcattgg ttcggtttat gtgatggatt acatttattg 132600
atttgcatat gttgaaccag ccttgcttct cagggatgaa gccgacttga tcgtgatgga 132660
taagcctttt catgtgctgc tggatttggt ttgacagtat tttattgagg gtttttgaat 132720
cgatgttcat cagggatatt ggcctggaat ttcctttttc agttgtgtct ctgccaggtt 132780
ttggaatcag gttgatgctg gcctcataaa atgagttagg gaggagtccc tctttttttt 132840
attgtttgga atagtttcag aaggaatggt accagctcct ctttgtacct ctgatagaat 132900
tcagctgtga atccgtctgg tcctgggctt tttttggttg gtggactatt aattactgcc 132960
tcagtttcag aacttgtcta ttcagggatt caatttcttc ctggtttaga cttgggaggg 133020
tgtatgtgtc aaggaattta tccatttctt ctagattttc tagtttattt gcatagaggt 133080
tttatagtat tctctgatgg tagtttgtat ttctgtggga tcagtggtga tatccctttt 133140
tttgtttttt attgtgtcta tttgattttt ccctcttttc ttctttatta gcctggctag 133200
cagcgtatat attttattaa tcttttcaga aaaccagctc ctggattcat tgattttttt 133260
gaagtgtttt tcgtgtctgt atatccttca gttttcctct gatcttagtt atttcttatc 133320
ttcctgctag ctgtttgctc ttgcttctct agttctttta attgtgatgt taggtgtcga 133380
ttttaaggga tctttccccg ctttctgatg tgggcattta gtgctataaa tttccctcta 133440
aacactgctt tagctgtgtt ccagagattc tggtacattg tctgtttatt ctcattggtt 133500
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
tcaaagaact ttattatttc tgccttaatt tcattattta cccagtagtc attcaggagc 133560
aggttgttca gtttccatgt agttgtgtgg ttttgagtga gtttcttaat cctgagttct 133620
aatttgattg cactgtggtc tgagagactg actgttgtga tttccgttct tttgcatttg 133680
ctgaggagtg ttttacttcc tattatgtgg tttattttag aataagtatg atgtggtgct 133740
gagaagaata catattctgt tgatttgggg tggagagttc tgtagatgtc tattaggtcc 133800
acttggtcca gagctgagtt caagtcctga atatccttgt taattttctg tctcattgat 133860
ctgtctaata ttggcggtag ggtgttaaag tcttccatta ttattgtgtg ggagtctaag 133920
tctctatgta agtctttaag aacttgtttt atgaatctgg gtgctcctat atttggtgca 133980
aatatattta ggatagttag ctcttcttgt tgcattgatc cctttaccat tatatgatgc 134040
ccttctttgt cttttttgat ctttgttggt ttaaagtctg ttttatcagg gactagaatt 134100
gcaacccctg cttttttgtt tttgtttttg tttttgtttt gttttgtttt gctttccatc 134160
atctgcttgg taaatactcc tcagcccttt attttgagcc tatgtgtgtc tttgcatgtg 134220
agatgggtct cctgaataga gcacactgat aggtcttgac tctttatcca atttggcagt 134280
ctgtgtcttt taactggggc atttagccca tttcatttta ggttaatatt gttatgtgtg 134340
aatttgatcc tgtcattatg atgctagctt gttatgttgc ccattagttg atgcagattt 134400
ttcatggtgt tgatggtctt tacaatttgg tatgttgttg cagtggctgg taccggtttt 134460
tcctttccat atttagtgct tccttcagga gttcttgtaa ggcaggcttg gtgatgacaa 134520
aatccctcag catttgcttg tctgtaaagg attttatttc tccttcactt atgaaagtta 134580
gtttggctgg atatgaaatt ctgggttaaa aattctttta agaatgttga atattggctc 134640
ccactctctt ccggtgagta gggttccctt tgtaggtaac ctgacctttt tctctggctg 134700
cccttaacat tttttccttc gtttgaacct tggagagtct gacaattttg tgtcctgggg 134760
ttgctggttg ctcttctcag ggagtatctt agcggtgttc tctgtatttc ctgaatttaa 134820
atgttgacct gtcttgccag gttggggaag ttttccctga taatatcctg aagtgtgttt 134880
ttcaacttgg ttcccttctc cccgtcaatg tcagggactc caatcaattg taagtttggt 134940
cttttcacat agtcccctat ttcttggagg atttgttcat tccttttcat tcttttttct 135000
ctaatcttgt cttcacacct atttcagtaa gttgatcttc agtctctgat accctttctt 135060
ctgcttgatt gattcagcta ttgatacttg tgtatgcttc agaaagttct cctgctgtgt 135120
ttttcagctc caacaggcca tttatattct tctctaaact agttattcta gttagcagtt 135180
cctgtaacct tttttcaaag ttcttagctt cctcgcattg ggttagaatg tgctccttta 135240
gctcagagga gtttgttatt acccaccttc tgaagcctac ttctgtcaat tcatcaaact 135300
cattgtctat ccagttttgt gcccttgctg gagagaagtt gcgatcattt gaaggataag 135360
aggcattctg gttttggaat tttcagcatt tttgtgctgg tttttccttg tcttcatgga 135420
tttatctacc tttaatcttt gaggctgatg acctttgaat ggggtttctg tgtgggagtc 135480
ctttttgttg acattgatgt tactgctgtc cgtttgttag tttttctcct aacagtcagg 135540
cccctcttct gcaggtctgc tgcagtttga tggcggtcgg ctccagagcc tatttgcctg 135600
ggtattacca gcagaggcta cagaacagta aagattcctg cctgctcctt cctctggaag 135660
gttcttcccc aacgggcacc agcctgatgc cagctggagt tctcctgtat gaggtgtctg 135720
tcaacccctg ttcggaggtc tctcccagtc aggagacatg ggagtcaggg acctacttga 135780
ggaggcagtc tgtcccttgg tagagettga gcgctgtgct aggagaacct tccttgtcag 135840
gatctgctgc tctcttcaga gcaggaaggc aggaacattt atgtctgctg aagctgtgcc 135900
caaagctgcc ctttccccca ggtgctctgt cctagggaga tgggagtttt atctataagc 135960
ccctaactgg ggctgctgcc tttctttcag agatgccctg cccagagaag aggaatctag 136020
agaggcagtc tggccacagc cactttgcca tgctgtgttg agctccaccc agtctgaact 136080
tccaggcctc cttagtgccg acagtgggaa aaaccgccta ctcaagcctt agtaatggcg 136140
gacgcccctg ccccccacca agctcgagtg tcccaggtgg acttcagaca gctgtgctgg 136200
cagtgagaat ttcaagccag tggttcttag cttcctgggc tccgtgggag tgggacctgc 136260
tgagcaagac cacttggctc cctggctgaa gcctttcaag ggtagtgaat ggttctgtct 136320
cactggggtt ccaggtgcca ctggggtatg aaaaaaacaa aaacaaaagc aaaaacaaag 136380
aatactccta cagctagctt ggtgtctgcc tgaacagttg cccaattttg tgcttgaaac 136440
ccagggccgt ggtggtgtag gcatacgaag gaatcccctg gtctgtggat tgcaaaaacc 136500
atgagaaaag cgtagtatct gggctggata gcaccatccc tcatggcttc ccttggatgg 136560
gaaagggagg cccccctcca ccgctccttg cactccccac tccccaggtg aggtgacacc 136620
ccaccctgct tctgcttgcc ctctgtgggc tgcacccact gcccagccag tcccagtgag 136680
atgaacagtg taccccagtt ggaaatgcag aaatccccca ccttttgtgt tgatcacgct 136740
gggagctgca gaccagagct gttcctattt ggccatcttg ccagatgccc tcaagactga 136800
tttttttaat catcactttt aaaatctaca aaacaggcta tggcagggga acctgaactg 136860
agtgtgttat ctgagcatat ctgaaaggca ggtggggtgt gcccggaagg actctacctg 136920
cagaaggtga gaggagcaac tagggagaga aagatgctga gaaaagaaag ccaaagagac 136980
aaatatggcc agtggcccaa atatggccat tggagatagt gaagtttctt tgcccacaga 137040
ttgtagttta tcctttttta ttttatttta ttttattgag atggagtctc actctgtcac 137100
41
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ccaggctgga gtgcagtggc acaatctcag ctcactgcaa cctccgcctc ccaggatcaa 137160
gggactctcc tgcctcagcc tcctgagtag ctgggactac aagcacgtgc caccacgccc 137220
agctaatttt tgtattttca gtagagacag ggtttcacca tgttggccag gctggtctca 137280
aactcctgac ctcaagtgat ccacccacct cagcctccca aagtgctggg tttacaggca 137340
tgagccactg cacccaggca acatatgtga atcttgaaga cattatggta agtgaaataa 137400
gccagtcaca aacagatgaa tactgtataa tgccacttac atgagctccc tagaataggc 137460
aaattcatag aggcagagag tagaatagag gttaccagag catgagggta gggcaggaat 137520
gatgagttat cgtttcatgg gtagagtttc agtttgggat gatggaaata gttctggagc 137580
tggatggtgg tgctggttac acaacaatgt aaatgtactt aatgccacag aattatacag 137640
taaaaaaaaa aaaggttaaa atgggaagtt tcatgttatg tatattttat cacaatgaaa 137700
acaaaataag cagttaagta acttgcccaa gacacagagc tagtaatgag cagaatgagg 137760
attaacagtc aggtgctcca gttcagagcc tgcactgtct accttttcca gagacacttc 137820
caaactggga ggtggcacca gctagcaggc atctgttggt ccttgctgga catcaaaggc 137880
ataaatatgt ggcaaaagat tctccaaaaa gagaaacaaa tgtccaccac acggtgattg 137940
ggagggcgac attcctgaac acacgacatg tttcctggtc atctccagaa ataatagagg 138000
aagagactag acagctcata gtaacatgtg ggaatgagtg actgatgtgg ttattctaat 138060
tctattttta aaatatacct ggtgtagcca gagacatact agctacaagg gcaaaacttt 138120
acagatgtgt gggtaccgga aaatatttgg aaatttatgg ttaatgcata tatatgaggg 138180
agtctctgaa cctattctgg ttcaggaggc tgcctgaaaa gaaaagaaaa tttaatatta 138240
aaaaaataca tctatatgaa gtccaagtac aatatgatga tttttttcat tacttgatgg 138300
cctgtacctt tcatgccttt ttaactgtgg agggtatatc aaaatgtaaa gtattctttt 138360
gtatattcca ttctacttaa ggtgattcaa acaggttaaa tttttcagtc tctggaattt 138420
agcaggattt agcagagtat atattataaa caaatatgta gaacaggtga tacatgctgg 138480
gagagtggtt gtctacatca ctgtaaatat accaaagaga aaggcattga tgccaagcat 138540
ggctgttgca agatacaagt cggggaagct ctaaaagtga catttgatgt ggcttctgca 138600
atcagtagga attttccagg cagacaacag gggtgaggag gggatattct aagcaaagga 138660
aagagcctgg gctgagacac aagggacatc acattgacag tgggtacagg caacagaaat 138720
taatgcagta ggactgaagc tgtgtttctc attgtgcaga cctgcataag aaccacttgg 138780
ggtgcttgtt tggatcccac acaagcagat ccctgggtcc cagcccagcc ctcctaagtg 138840
ggacatttct agtaacagag gaggggaata cattttcaac gatatcctcc tcgtgattct 138900
tagcacacaa aagtgcagtg gaggggatca ctgagctgga ctggggagcc cctggttgga 138960
aatgagtgtg gagaagcagg ctggggcaag gttgcaaagg atcttgtact ataggcagtt 139020
gtatttggaa ggtagacaga aaggagataa cctgattgtg ggcagattct ggaaaaggtg 139080
ggagggggaa gggaaagtgc agctgcagtg aagcaagttt tcttattgag aggtgagttg 139140
tgatggtgga tgagggctga gagacaatga atactggtga tactggattc acggggctcg 139200
atgatgcgtg agaactgagc gtggagtcca ggaggcagga gcttcgagag acctgaagag 139260
aacactgatt cccagcattc tcactgcagg tcttgtggaa cccagtgtgg gcccgtcccg 139320
ctgcatggct gcagctttat tggtgtaaac ttgtagggac atgtggttag tgccaggtgc 139380
ttttattaag gatgctgttt ccctattcct gaggactcac gggaattttg.agaaccaact 139440
tctcccattt ttatgagaca agagcgcaaa cgctttagga aatatatggt tattactttt 139500
ctgtggtaat aaccatttat aagctgcggg tgttttcaga aagtatttga tggagaatgc 139560
tacactctta ctaaaacccc aaggggaaca tcacttcttt actcttttta ctctttcatt 139620
gtgtgtcacc tttacttgtc aacttcaggg tacataaagt tgcttccagg tgcataaaaa 139680
gagtatggga aacccctata gtttaacttt ataacaaaaa agtatattca aaagagctta 139740
taattgcata gcaaaaacta acacaaaaac aaaacacatt catgcaggaa taaaaagggg 139800
tctcctaaaa gtcggttttc acacctctgt tagaaaacga agtttcgaag tatcctcaag 139860
cttttggtct attacctgtg agtacctcac ctatgacatg gatggacact gtgactagtt 139920
cacagcaaga tttctcctcc taggcactgt tgacatttta ggctggatac ttcttgtttg 139980
aaggtggtga gcctgtcctg tgtattataa gatgtttagt agcatccatg gcctctacct 140040
acaaaatgcc agtagtaacc cttcttccca aggtcataac aattaaaaat gtctccaggc 140100
tttaccaaat gtctcctggg gtatagttct cagatgtata aaataaaaat acaggaccct 140160
tagttacgtt tgaattccaa ataatggaca aactttttag tagaagtatg tcccaaatat 140220
tgcattggac atacttacac taaaaacgaa tttttatcag atgtaactgc tggcctgtat 140280
tttatttgat tgccctacct gggcagccaa caagatcgtt gatgtacgaa acaggatgcg 140340
tgtgaggtgc ctggcacatg gtgggtgcat tataaatatt caaggaatct gctgcgtacc 140400
tttgaatcca ctttacccgt gtccctgctg tagtctgcaa ttggtactcg agcctgccta 140460
tcctttgcca aaactgtcat gagagtgggt gtgaggagat agaaagccca catttgaatc 140520
tgcaagttct gggaactagc attctggtca aagctggttt tacatccacc tctgccagag 140580
ggagcatcaa tctgcctatg ctcgagcact ttgtcactgt ggtctccctg agtcattaga 140640
tcatttctgg taagtagcat tcagagaagt gatagtgaat gaaaaactgc tgcattagtc 140700
42
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ccagactcag tcactctgca tttgattttt ccagcagcat agccctagaa aaccgttcac 140760
tgggatgcag caggaccaat catttccaca ttcagttgca gctaggaact tatttctgag 140820
ccaagatgaa tgacccaaat agcctttcca gagagcagag gtgaaattta actctgggtg 140880
gttgcctgac attttgagac aaaggagaaa ggggatgggg aatgaagaaa aatagcattt 140940
gaatgtctcc tgcagggtca taaatctgga tgggcaaaag tggattgcat ctctgccaac 141000
aatttaatta atttgggctt cttttaatct ctatgattca tataacctga aggaaacatg 141060
ctttaaagct gtattgtagg aagagcctcc tttgagttac atatcagtat attaactctt 141120
ggatgtttta tttttccaga aggaagtcta agttgtgtgt gggggaagta ctaggtttgt 141180
tcacaaaaaa atgcattcat tgctttgggg gaaagccccg ttcctgtctg actgtagtga 141240
gatgaatcat tagaagtgta atcacagccc aggggaggaa aaacaagagt aaaggattct 141300
gatggctccc agcaaagcct gagagtgggc tggagatttg catttgctta ttcttgctgg 141360
tgagcctttg ctgatcatct ccaacttata aaaagttgca agtctatggg aaactgtttt 141420
gaaacaaggt aatacaaaat ggcagctatg acactacaga atagcttggg gacatatcta 141480
gctgggatgt gtattgcgtt ggggcaggat gtatattgtg ttggggctca aacgagcaga 141540
tgcgaagcct tttatggctt tgctactcaa cagaaaggtg tttcattttt taatccctat 142600
gatttattta tttatttatt tttattattt ttttttttga gacagggtct cactctgtca 141660
cccagggtag ggtgcagtgg tgcgatctca gctcactgca gtctctgcct cccgggttaa 141720
aacacttctc ctgcctcagc ctccctagta gctgggatta caggcaccaa ccactgcatc 141780
cagctaattt ttgtattttt agtagagacg gggtttcacc atgttggcca cgctggtctc 141840
gaactgctga cctcaggtga tctgcctgcc ttggcctccc acagtgctgg tattaccagt 141900
gtgagacatc acgtctgacc taatcgctat gatttttaaa agtgaaatta tcaggcaaga 141960
agaaatgtca aaagggaaaa aaaagaaata aaaattaaac gattccattt tgtgtcttca 142020
gtggtaattc aagtaaccat gattttctat gtttcttcct cctcaatccc acagtttggg 142080
gctcaccaat tccttaacat acgctctgcc aaatattagg gtgccaggaa gtatattggc 142140
aaaggaatat agatgtgtct tgattttata acaggtatat tgttccattt tcagtaacgt 142200
gatgggctat gttattggga ctacccacaa aaaacaaccg aaaagagttt aacatgacaa 142260
ataggaggaa aaaatcagtg agtgtataga tgatttgaac aacacaatca acaaacttga 142320
cctaatagat gtatgtagac actgcaccca atacctgtag ggtacacaat cttttggagc 142380
atacgtggag catttataaa aatggactat atactgtgct ataaagcaag ctccagtgta 142440
tttcaaaaaa actactaagc atgtgttctt tttttttttt tttttttttt ttgtattttt 142500
agtagagaca gggtttcgcc atgttggtca ggctagtctt gaactcctga cctcctgatc 142560
tgcccgcctc agcctcccaa agtgctggga ttacaggcgt gagccatcgc gcccggcgta 142620
tgtgctctta tcacagtgaa attaagccaa gaaatcagct ttttgaaaat taggaaacat 7.42680
aataaatatg tttctaaaaa ttccagtagt cacattttca tccactgtac ttggagggag 142740
agtatgatgt ccccttgctt gaggcacaag gaaccagaaa ttagagctcc cagtcattgt 142800
agttgcatgc atttccccac ccattagaat ggaatttctt tgacaaaaat gacaaaactc 142860
taagggtcat tttgggcctg agtcatgagc taagcagctg tttcaattgt caagtgttaa 142920
atgatttgtt ctggatcaga gactataagg caagaaatgg ggctaggaaa aagaaacagg 142980
ttacagaggt caacaagtta gcagaggtat acaagtgtgc cctttgggca tcagcaacaa 143040
agagcacctg ggctgacctc tgctgtctta atgctggaaa tgttatagcc cgctctgtaa 143100
tctctccccc ttgcccccgg gagaaaaatc tttcacctaa caatggattc cactagaatt 143160
tttgtattgt tatcacaaaa atcttctagg aataaacaga gagcttttgt tgatcatgag 143220
caagatcttc ctggggccat gtgcaaacag caaaaagaaa atttgatctg gaagagaatg 143280
agaaccatgc agttgtttgg aatactattg ttgggctttt tgggttttta cggacagcta 143340
ctttgaaaag cagattgact aagttgctaa tatgtgaaat aagcaccact attttgctgc 143400
aagtcctcta tcaacttttg gacacttgtc tcaggactaa ctgcagttta gaaggtgttt 143460
tgttttgttt tgttttttgt tttgaaagta tttcgtttct tgccttagat ctatacttga 143520
aaggaccttc ttgtggcctt atctggtaga gacacaagat cttagaatca gagagaactt 143580
tagttctgga atgagcttta gaggctagta ttccaacttt ctacccaatg taagaattgc 143640
caatatttac ttttgtttgt ttttcagtct cctaagactg tatgtgttca taaaatcagc 143700
ataattgatt tgttattatt catccacaga catcagctaa tataatagtt tatatataaa 143760
tacaaaagta tacatttcac tttaaataaa catttatgtg agtaaaatag agaaggaaac 143820
ttaatgtgtg cctatttgtg ctttaagata ataacatgaa tatatacaat gtctatttat 143880
agactaatga gctgacttaa catgcattta tgaatactca agtcttcagt tgcatttggt 143940
caacatttga ctatcctaca ggctctttta gtataaattt gcataaaaat cttttacaaa 144000
aaagatatat atatatatat acacatatat atatttttaa cttcaggctg attgacctat 144060
actatcataa cattaatact atattattat ctatatacaa aagaaaatcc caaaggacct 144120
ttgcttaaaa tatagacaca cacacataca catgcacatt tcatgctgtc agcttttttc 144180
aactctcaga ctcatcattt ctctcactgg agtttgggaa gcaccatgct tgagtcagtc 144240
ttacatgttg gccccaggaa ccctttctga aaagttcccc tcattcactc tttgcatatt 144300
43
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
tggggcactc acagaagata ttccagaaaa aaaaatgacc ctaaaaaaat aagcccatga 144360
atttgtgagt ttcaaaagcc accagaaaat gtgcttcttc gaaagcacgg ttagggtcat 144420
aagtcaaatt caggaagcag aggcataatg tgacttcctt ccaccgctgg ctgctcctct 144480
gcgcattcca aagggcaaga ccagagcttg gaaacagaaa tagaaaaact cgcaggaaag 144540
gttggcttgt tgctcgggcc tgctcttaaa gacagatgac tattggagtt tgttggaatt 144600
taacccctgg actctctccc tgggatgtat aattggctct cacttcctga atgcatgatt 144660
atgtgctgca ggcagatcgc tgccaattat ttttattgtg attgctttgt gcatctctgt 144720
tggcaatagc gttcacagga tttcagctca aggggaacgg ccacagcccc aagggtccaa 144780
gattagaata tgactgcggt tataggtcaa aagattaaag gtctgtgaat attgttctct 144840
ttttcaccca gagataccag gatagaatct ggcactttcg ccttggtatc cataagtgag 144900
acctgtattg tcagtgttta gatttgtcac aatcagaatt aacattcgtt tagttcagaa 144960
aaatatgtaa gtattgaata gtggcacaat tataatttga ttctttctcc tctctcttct 145020
cccatcaggg cctgtgtctc aaaaaatgta aaggcttgca actctttgaa aagctttctc 145080
ttttcctttt tgggtaaaaa gttatacaag acactagaaa attcaaacat taaagaaatg 145140
tcaagtgaaa atcttcagaa tggaggacta taattttttc aaaaagtaaa agtcctcatg 145200
agcttctcct ccctaccctg agtaaccact ttaaatagtt tagtgggttt tcctcctagt 145260
cattttctag atatgtttta tgtatatata taaaaactga tatgttttat gtatatatat 145320
aaatgaaatc actattattt aaaagtgctt ttttcactga gcaatgtcac ttggacattt 145380
tccatatatc tgtttagaaa aatcgacctc attctttccc aatagttata tgatattcca 145440
ttgtaaggct ataccatagt ttatttaact atctcccctc catgtgcccc cctccaccta 145500
ctacttggcc ttttttttcc tttttctcct tctttttttt tttttctttt tacaaattga 145560
gaaggagatg aaaaatctct gctggtatta gcaacaggaa ccatccagct gcatctggtt 145620
aaatctggtg gagtcacaga tgcatgacaa cctgagtttc cagccaagga ttaatgtttc 145680
atgcagatta catagtattt atattttaaa cctgtgacat gttgagggga taaaatttta 145740
tatctgtagg aacaaaatca gaaatttatc tgatcattgt cctcttccta agcttgttgg 145800
gcccttgaaa aaatggattt tgggcccctt tttaaagaaa attttagttg gccaggtgcc 145860
ttgtgagtaa agaagaccat agacaaatca atacaggtga tttacttctt agcttaaaaa 145920
ttgatctcac atatgatact taatgttttc gtttagaacc ttgggtttgt gtcttgtacc 145980
taaaatctgc attcctacca aaaactattt cattaataat atcacataac cgacagtcaa 146040
atacagatca gtaggccagg tgcagtggct cacgcctgta atccccgcac tttgggaggc 146100
cgaggcgggc agatcacgag gtcaagagat caagaccatc ctggctaaca cggtgaaacc 146160
ccatctctac taaaaataca aaaaattagc tgggcatggt ggcgggcgcc tgtagtccca 146220
actactcggg aggctgaggt aggagaatgg cgtgaacccg ggaggcagag cttgcagtga 146280
gccaagatcg tgccactgca ctccagcctg ggcaacagag cgaaagtctg tctcaaaaaa 146340
aaaaaaaacg aacacaaaca aaaacaaaac aaatatagat cagcaaagta ggtgtcaagt 146400
ccccagggga tcagggacag agcccagaca agaacatgaa agccagcatc cttgtatgtg 146460
tgtcacatac ctctgccttg gcaccttgaa aggtttcaga cagtcaggca actggggagg 146520
cagctgtttt ctgcggtcac agagcacatc agcttaaggc tggtaaaaag ctttgattat 146580
tattcatgac atataatact tttattagct ccacactgat tagtgctgtg ctagaactgc 146640
tttctgtaaa attgagatcc ttccttgcat ggaacttata ttatctgtag ataaattcct 146700
cggtatgaca tttgaatgtc ttaacatttg agtatcttaa caaagtccaa aggataatta 146760
gaatcatcac tcatacaaga catattttga agacaaaaag gtgtggtgcc aatataaaag 146820
gttcatttaa aagaatccat tttactaact gcaggtaagt atccaaaagc aacaacccac 146880
tttcccagtt gattgtggct tgttttccca ttcctgtcct tattccacgt ttgctgtctc 146940
tccactttct gattttgtag tgtaggcttt gtgtagctct caaattcaaa tttaagccct 147000
taaaatggag ctcaaaagca ggtcctgaga aattgatggc tgagtagtga tggttgtcag 147060
tgttttacac cttcgttctt atatctatgt ttaacccaga gaatggtatc atagttttac 147120
aaagcatttg cagggtcatt tcttgaggcc ctcatggtgt ctatctctca ttcaagccac 147180
aggttatcac ccatcagcct ttgaccctgc cccctgctat gggacatgcc acaaggcagg 147240
aggagtaccc aacacccttt ccaggactcc ctctcagggc tcaagtgatc ctcccacctc 147300
agcctcccaa gcagctggga ctacaggcat gtgcctggct aacttttgta tttcttgtag 147360
agacgaggtt ttgccacttt gtgcaggctg gtctcgaact cctgacctca ggtgatccac 147420
ccacttcagc ctcccaaagt gctggaatta caggcatgag ctacagtacc tggccttttc 147480
tttttctttt tttttttttt ttgagacagc atctcactct gtcacctagt agctatgacc 147540
acaggtatgc accaccacaa ctagcttttt tcttcctttc tttcttttct ttttttttta 147600
gagatgaagc cttgctatgt tacccagtct ggtctcaaaa tcctgtcctc aagcagtcct 147660
gctgccttgg cctcccaaag ggttgggatt acaggtgtga gccaacacat ctggccttgt 147720
ctgttaacat atctgccttt ttcaatggat tataacttcc ttgagatcag ggaatcagtc 147780
tccatatcac agcattatga caggcacata taggcactaa agtaggaatg aatgaacaaa 247840
aagtaaatga agatatcact aggcattagg gaaacacaaa ttaaaattgt gatgagatac 147900
44
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
cactagatac ctataagaaa ggctaaaatt taagaaaaga aacctaacaa gtgaagggaa 147960
agatgcgaag cagctagagc ttgcatacac tactggtggg aattcaaaat ggcacaacca 148020
gtgtgcaaaa tggtttggca gtctgctgta aagctaaaca tgagcttatt atgtgactcg 148080
accattccac tcctacatat ttacccaagg aaattttggg ggcacacaga aacctgtatg 148140
aaatgtatat agcaatttta ttcataatca cccaaaactg gaaacaatct aagtgtcctt 148200
caactggtga gtggataaac taaatgtgat atatttacac aacggaatac tacaacacaa 148260
tgaaaaagaa caaaccacag gtatgtgcaa cagtttgcat aaatctcgac tgtaacgcaa 148320
agtgtaagaa ccagaccaag gctgggcaca atggctcaca cctgtaacct cagcattttg 148380
agaggccgag gcagaaggat ggcttgagcc caggagttca agaccagttt ggataatatg 148440
gtgaaactcc gtctttacaa aaagttagcc gggcatggtg gcgcttgccc gtggtcccag 148500
ccatttggga ggctgaggca ggagaatcac ttgggaagcc aaggctgcag tgctgagatc 148560
acgccactgc acgccagcct gggtgaccta ggagacctgt ctcaaaaaaa aaaaaaacag 148620
attccaaaag ctatgtacaa tatgattcca tttatatgat attctagaaa agacaaaact 148680
atagggacag acaagagatc agtgattgcc aggggttagg gctggaggaa gaggtgactc 148740
tcaaggggca gcaggaggga atgttttgag gtaatgaaac tgttctgtgc cctgataatg 148800
atggtggtta catgactctg catttgtcaa gctcatagaa ctgtataact aaaaacaaat 148860
aaagttttac tctatgaaga taatacaaac ttaatttgaa aaagtgaatc aaggttattt 148920
tgctgtttag tgtagattca gaaagaaaat aatcaaaaga aaatctgggt ctgtggtcct 148980
gtaacctgca gtgattttgc ccatgctcat gtacacattg tgtgtgtgtg tgtgtgttca 149040
acaaacatta aaagaacact tcctatggac tagcacttgc ttaggaggtc ttgatgagtg 149100
ggaaaagcag ggttcctgct ctcttggtgg gagagggaca ggtaatatgt aaacaacaca 149160
tgaatatgac aacttcagat agtgataaga gctgtcaaga caataaaaca ggcctacgtg 149220
agtgggattt tgtgtgtgag cctctctgag gagacaccct taagcgaatc ctggaaggta 149280
cgaagcctgc agccatgcat gcaaatactc aagtagagct gtccagacca ggaaaacatc 149340
acattccaag actcctaggg gacaacattg aaaaccctca cttcaaaact cagaaagcct 149400
ggctaaaata caacaaacat cctttctatc ctttctcatg cgtggctggg cttccaggaa 149460
aaaggcttag cccccaggat gggattaagc tctatgttca gagaactgga agcaggtcca 149520
tgggactggc tcaagccagt tagaggacca gcagaagatg agctcagata catgaatagg 149580
aagcagtcta cataggcctt aggtgccacg atgagacatt tgggtttcat tccaaatgta 149640
ctcagagctg tgggaggctt tcaagtgaca gagtgaagtg accttattga cattttaaaa 149700
ggataactct ggctgctata ggaagagtgg actatggagg accaagagtg acagggagac 149760
caaacaggag gctgttgcta aaggtcaggg ggaagggcca cgtcggccag aacctgtgca 149820
gtgccatgga gatagggaga agtgagtggc tttgaggtag attttagagc tggaacttgc 149880
tgatggattg aacattgggg attgtgaggg aaaaagaaga tgaggatgag gtctcagttt 149940
ttgacacgtg tatctggggt ctgacagtgc tgaaataggg aagggtaaga gagaaacaag 150000
agttttcctt gatagtgtgg caggaaacca aaattccatt ttagacatgg taactttgag 150060
acacccatta ggcatctctg tgaagatgta agtaggcagg tcgaggtata tttctggagc 150120
tctgggaaca gaaaaagaat gtatttcaga gacagtggca tgcaggtggc acttacagtc 150180
ttggagatag attagtgccc etagaaaggg cctgtggaga gaggagatga ctaatgacca 150240
agatttggac tgtgccaacc tttgggggcc agtcagagga gccagctaag gaaactgaag 150300
gaaagccagt gaggtgggaa cgaaactagc atgtagtgat gcagaagcca agggaaataa 150360
gttttacaaa aaacagtgga atggacagct atacagaatg atttagagag tttgattaga 150420
gtgcaaccat taaatatgac aaagactggt ggccttggtt agcgcagttt ctgtggaatt 150480
tgggaacagc ctgtttgggg tgagaatact gggggagaaa gtggagactg gctatagacc 150540
atccttttga ggttttactg taaaggggaa ctgaggaata gcagtgcagc tggagaagcc 150600
tgtgggagtg gaggaagggc tctttctcag gtgtcagata gagatatgct gatagaatga 150660
cccatcaagt aaagcgagag aacatgacga tgagggttgg agaggaggaa cagatactta 150720
caggagttcc gtccttgagg gaaagagaag gatgagctcc accgcccatg tgagcagggt 150780
ggccttagct aggcacatgg acggctcatc agttgtaatc aaagagaaga cagggagatg 150840
gaacaggggc aggtggatga atagattcaa ggatgagaag atgaggagta tgtagttctc 150900
tctgtgttta tttcctctaa gaaataggaa gtaaggtcaa gtgccaggag tggagttggt 150960
tgctggagat ttgaagagaa atggaaaaag gtataaaatg gttttggata ctggggaaaa 151020
aaagcatatt acaaacaaat ggagagagaa atgagtggac aacacaccct tagagagaga 151080
gagacaccac cagcatgtnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 151140
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnc 151200
cctggcccag acaggctcat cctgccaagc ctaggtctaa gcacaatctc agcttttgga 151260
gccagcagtt gcagccaggc cgaattcagt ttttctggaa ttcagttgtt ctgacccaag 151320
ttcagccggc aagtgagggt ccgctgtgtt cctggtgctg agagcgcgac agggtctgag 151380
cctgccagga tgacaggatc ctggtgatgc ggcccaacag gaagcaaagc acagggaggc 151440
agaaatgaga aacagtatct tagcccaagt tctaaatcag tcggcccctg gccaggttaa 151500
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
gtaacttagc acgtgtaaag actaaaaaaa ctaaagcagt agagaattac cttatacaga 151560
tgacaagata tggacaacta agtgagaagg tatcacaact aggtttaata gaaatcctta 151620
aaaaaagtaa gccgacaaac agaaaagaca acaacagtga aattcaacag aagaaaagta 151680
atagactctg atgaagatga cgattattga actaaaagtt ttcatagact agaacttaat 151740
ggaacgattc taggatggaa gttaagatct gatttaaaat ttactttgtt tattgtctat 151800
atgccttttt taaaaataaa cttgttatgc aaagtaaaaa aaaaaaaaaa ggcagggagc 151860
ctgaagtttt atagagaatg ctacggttct ccaacttaac ttctatcagc caagttagca 151920
gtagagcttt aactgtaaga agaagtgtat tggtttctgt tgctgctgta acaaatcgct 151980
ataagcatgt tggctttaaa caacacaaat ttgttatctg acagttctag agatcaaaag 152040
tttaaacgga ttcgcagggc ttctagagtc tccttctgga ggctctagga gagattatgt 152100
gttcttgtct ttccagcttc taccagccac ctgcattccc tagtttgtga cccactcctc 152160
tatcttcaaa gccagtagca cagcatcttc aagtctctct ccaaactaat ctctgcttcc 152220
CtCaaCCCat CtCCCtttCt cattgtccct gCCtCCCtCt tttCCttCta gggacatttg 152280
tatttacatt gagcccaacc atataatata aaataatctc tttgtctcaa gatccttaac 152340
ttaataacac ttacaaagcc ccttggcatg taaggtaaca tattcacaga ttaggatgag 152400
gacatcttta ggggtgaggg aatattctac cataagaagc ttcctgctgt atccattgta 152460
gtgactttct ctttggttaa atcacttcat cctatttgcc tttgaaaact taccttctga 152520
actaattgag gttctgtgtg agaaacatgt gaggggagaa gaaaaggcac acacacaata 152580
cctttaaggg taaacaagct ttatcccacg taaatggcaa tgcagatata ataagcaaat 152640
tgatataata agcaaatgat ataataagca aattgatata ataagcagat.tgacataata 152700
agcaaattgc aatgggaagg agagaaggga aaagagattt acattcacca gactatggag 152760
gattcacccc cagactggga agtaacagcc tggactccaa agtcagccac tcatccatgt 152820
acagacaagg agaggtctca tgaagctttg gcgcagtctg ggaccccagc tctttttgta 152880
acaaattgtt tggcatgagg cccagtcatg agggcccttc tcgactaggc tcaaggaaca 152940
caaaaaagtc aacctgtttt tgtgattgtc tattgttttt caataactaa tgtgtaagag 153000
tcgattgaaa tagagatttc tctgaaacag tgctggatga atgcctcaag tggctcacat 153060
aacctgttct gggacttggt gaccattgtt tgtgtccatg ttcatctgag ttcaaattta 153120
atatttaact tttcctccac aattgattag aacagtggta ccagaggcta ggcacagtgg 153180
ctcacacttg taatcccagc actttgggag gccgaggcag gcggatcact tgaggtcagg 153240
agttcgagac cagtctggcc aacatgatga aaccccgtct ctactaaaaa ttcaaaaaaa 153300
ctagccaggc atggtggcag gcacctgtaa acccagctac tcaggaggct aaggcatgag 153360
aatcatgtga acccaagagg caaagattac agtgagccga gatcatgcca ctgccctcta 153420
gcctgagcga cagagtgaga ctccatctca acaaaacaaa acaaaaacaa gtctgggcgc 153480
ggtggctcag gcctgtaatc ctagcacttt ggaaggctga ggtgggcaga tcacgagatc 153540
aggagatcga gaccatcctg gccaacatgg tgaaacccag tctctactaa aaatacaaaa 153600
attagctggg catgatggtg tgcacctgta gtcctagcta ctcgggactc ctgctgaggc 153660
aggagaattg cttgaaccca ggaggtggag gttgcagtga gctgagattg taccactgca 153720
ctccagcctg gcgacagagc gagactccat ctcaaaacaa aaaccaaaac aaaagcaaaa 153780
gaacaatggt accagagacc ttcccttctc ttgccttcgt tgtttccccc tatactttat 153840
gaaatgtgct gtggcctgaa atttaaaaca ttgcatttgc ttaactcagc cacctaaagc 153900
ttttattttt ctttcagtgc cttaaagtaa taggatttag agcttttaaa aagaagttct 153960
ttatatgaag ctctgagatc aaatgtgaag tcaacaagtt gttttctctg atccatcata 154020
gggttttttt ctgcattttg gatatgtgag agttttgaac ttctgctaca taaaacatca 154080
tagcaatcat acaccctaca gtttatgata taattaatat ggtaatatac actatcttca 154140
tatattaaat tcccagagtc tactaatcta agattaatat agcatgtcat tgatgtttac 154200
atcatttgtc aacaaattca gcatttaaga atgagtaagc atgcacacac atgaggctgt 154260
gtgtggctgt aagattctgt ctcaagtcct ggcaggaaag actaaggtgt aacttcagct 154320
ctgtggcctt gaggaagtta catcaccttc agaatctcac ttggctccca ggtagacaat 154380
gagtaattat acagctctcc tggagatagg atgtatatta tctgattttt gttcacaaaa 154440
aggcatataa ataccaagca ttgctataaa ttaaaatttc acctcacata attcatctat 154500
gcctttataa atctcctctc tgttttcaat tagatttgat atgcctcatt atttctcatt 154560
tagaaccagc cattggaatg aattttatat tcatttgaat ggatataaaa tggtttgctt 154620
tcaaccattt tatgtttatt caaatattag ctttctttta gttcttgaaa ggatttctgt 154680
ttatatccca ttatgcttat ttttctatta tataaaaatt tatattttta taatatttag 154740
aagttgaagt gttctattta gaaatgtaaa attctctgga tctatttcat gaggtggatg 154800
aggagacatg cagcaaccgc caattcctaa taattgtact ggtcagggtg ccagcaggaa 154860
gctagtgaca ccctcaaaag gggccgtgga aaggtgttta gcgaagggac tctttatgaa 154920
gatgtgggaa ccattcagag aaagcaataa ggaataataa agcaccctgg atccacctgt 154980
gcctcaaaga gcaacaggag ggaggggtgg ccagaccctg aaaagagcgg tggctgctgt 155040
gaaagaaggc ccaggaaatg agagcacaca ctgccagacc tccagccagt cagggggaag 155100
46
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ggagaaggga agcaagggaa ataatatccc aaccctcctt cactctgttc tcttgccttc 155160
cttcctttat tcaaacttag ctacaagcca gtgggcaagg gagccggttg atgcagtcca 155220
cagagggcca ttccctagca cacagcagag tgggaaagag tggagagtga tctggaagag 155280
caaacaaaag ataactagca cagtggtcta ttggaaaaag catcccaaat ttatgttggc 155340
ctttggggtt tggtacatgc atacctgcac ccccatactt aggccttgac ccctttggtt 155400
ccaagcacct ttcttctcag ctctgccatt atgggtctta aagctgagct ggccaccatg 155460
gatgatgccc agatcagctc actgctctgt ggatagtaag acttgactaa taagtccatg 155520
gttcgggcat cccgggtgat gttcccagga ggctgtctgg gaggagctct ccctcactct 155580
ccgtgtcctc cttttccagg gtggagcttc agcatgggat cggcctacca gtttcacagt 155640
tggcgtgtgt ttgtcatcgt ctgtgcactc ccctgtgtct cctccgtggt ggccctcaca 155700
ttcatgcctg aaagcccacg attcttgttg gaggtaacac ttattattgc agatactcag 155760
gtagcccacc tctattggaa aaagttcctt gttaattcta ggaaagcaca tttgcctatt 155820
tgagaggtac attttccctt cctattatag aatgatggag ggtttggttt catggagacc 155880
tgctttcaac aaccaacatg taaaaaagct gcctcatggc tagaaagcac tcatgtttct 155940
ccttaaacaa ataacttggc tcaatgaaga aaacatgatt tattgtttta tagcgaaatt 156000
ttaccatctg gagatactat ttctagttat aaccatatta tttgaactgc tcatgtattc 156060
agtgatatcc aaatagtctg tatgttccaa tgggatgtac tgagatacat ttgcacaatt 156120
acacctcatt tgatctgaca atgcaagacg ggctattgga aaatcaggga gggtaagtgt 156180
gtcactccta gcgcttcact gtccactctc atttcttagg ttggaaaaca tgatgaagct 156240
tggatgattc tgaagttaat tcatgacacc aacatgagag cccggggtca gcctgagaag 156300
gtcttcacgg tgagtcttct ccccagagaa tcctcaacac cagggattgg gacatgtttt 156360
ctgtcaaggg aaaaattgta aatattttag actttgcaag tcatacggtc tttgacacag 156420
ctactcagct ctgccattgt agtgcaaaag cagccatagg caatgttaaa ataaatggga 156480
tgactgtgtg tcaataaaac tttatttaca gagacaggca gagggctgtt tttggcccat 156540
aggttgccaa ccttgcttat gcctatctag gactcgaggg ctctgtggtt ggttaaatga 156600
taacaccaac ccaagagatg gggttaaggt ggaaagactg cattttcatc acagacttac 156660
ttactctctg actcccaggt tcagtacccg aggacaagag attcttggtg tagggactgc 156720
cagctgattc tccttgacct gctaccagaa gagagccagg ctgtgcaggg ccatttcctc 156780
cactcggcca ggtcatacct tttcaagtga cactcagccc ccacaaaggt gtatactcat 156840
agcagcccca taacattttc tctaagatgt cagagagaca tcttagagag gtatgcctct 156900
gttaggacca ccatgaggcc tttaacagac cttggaatac atagtaaatg gtaaaattat 156960
ttattgaaaa caaaaaaaaa ccagtatcta ataagtgaga aaacaattat actaaggatc 157020
aaagcaatgt aacaacataa aactaatctc tgggtttcct gatgtattag tccattttca 157080
tgctgctaat aaagacatac ccaagactgg gcaatttata aaggaaaaag gtttaattga 157140
ctcacagttc agcatggctg gagaggactc aggaaacttc tgatcatggc agaaggggaa 157200
gcaaacacgt ccttcttcac atggcggcaa caaggagaaa tgctgagcaa aagggagaaa 157260
atccccttgt aaaaccatca gataccgtca gaactcactg actgtcatga gaagagcagc 157320
ataggggtaa ctgtccccat gattcaatta cctcccactg ggtccctccc acaacacgtg 157380
gggattatgg gaactacaat tcgagatgag atctgggtgg ggacacggcc aaaccatatc 157440
acctgacaac caaagcaaag gataaattta atgaattata tcattattga tataaagcaa 157500
tgtgttcgct tttcaggaaa tgcttttcag gtagtgtatt cacttttcag gaaaggccaa 157560
ttttatcttg ttctaatttg tgaaaagttt attcataggc cttattataa gggatattga 157620
acaatataat ggacattatt ttcagtaaca cttttacata aaggcagaca tttcacacat 157680
ggccatttct taggttaata attaatcaga atttggtaaa ggaaagtagc tatataggaa 157740
accagagaaa ggtaatccaa gaatgtggct tcaacttgat atagcttgac atgaacataa 157800
ggtataaaag aatcatagat atagttccaa aaatatatat ttcattttat tatacatgca 157860
ggcacatggg cacacacata cataacacat accccagttt cacattctag cagcttccca 157920
ttgctctcag gataaagaca gaaaccctCC ctccggacac taaggtcctg ccagtctcct 157980
tcctgcctgc tttatagttc ccctctctct gcagctacat tcatccctca gctcctcaca 158040
ctcaacattt gacttactat acagcactac cacagtgctg ctgttccctc tgcatggaac 158100
attcctccat ctctagttac aaagctgatt ctccatgggg aaagccagta taaattaatt 158160
taaattggtt gcaggaaatg tttcggaagg aacaattgtt taaaatcttt eaatatacgc 158220
agtcatataa ttagactaaa aatggccagg cgtggtgact cacacccata atcccagcac 158280
tttgggaggc tgaggtgggc ggattacaag gtcaggagat cgagaccatc ctggccaaca 158340
aggtgaaacc ccatctctgc taaaaacaca aaaatcagct gggtgtgatg gcacctgcct 158400
gtaatcccag ctactcggga agctgaggca ggagaatggc ttgagcccag gaggcggaga 158460
ttgcagtgag ccaagatcgt gcctctgcac tctagcctgg caacagagca cgactccatc 158520
tcaaaaaaaa aaaaaaaaaa atagactaaa aatgttttgt ctacagtagg ttggagggac 158580
atatggtaag acataagtgg gaacctaaat aagatcttca gtggcttata cctaacaaac 158640
atcaattaca gttcttaaaa cacatgaaat cacttctgat taaatagtcc cctgctatct 158700
47
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
catttattaa tttcatttta tttaggaaat gaatttttca tactgtaaag ataaactgca 158760
atttggcctc tatccaaatt atcacagatt cttaattagc aaaattttct ctgcagtata 158820
atggagaaat gaagataagt aatggatatc ctcaattaag gcctggaagc ttttgaggac 158880
tctcttcttt tgagtagata tacaatagag gggaaaaatt ttaatttaaa ggatcataag 158940
gctggaagaa tcttgagaca ttatctggat atacaacatc acttaactct tctcagccag 159000
atgaagaaaa accctttatt taccagacct tctgaggagg tgtctctata gggcctcatt 159060
gccctactta cctgcctttt cttccaaaag ggttgagtta ccaagtccac taatggcgat 159120
gtgttaatgt tgttacatga aggttagagg agaataagat ggatatactc caaaagtgat 159180
tgataatttc aaagtcattt atatgtgact ttcaaaaagt tgttacatga ttctgccata 159240
acaaatttat ctttttattt tttaaatcca cggtacagca aagttgcatg cattttccta 159300
aggtctcctt ttcttttttt tttttcctta ttttttgaga cagggtctcg ttctgttgcc 159360
caggctggag tgcagtggtg tgatcacagc tcactgcagc ttcaacctcc ctggctcagg 159420
tgatcctcct gcctcagcct tcctactaac tgggaataca ggcatgtgcc accatatctg 159480
gctgattttt gaattatttg tagataatta acaacaggtc tccttatgtt gtccaggctg 159540
gtctcaaact cctaggctca agtgatcctt ctgcctcagc ctcgcaaagg gctggaattg 159600
cagatgtgag tcaccatgcc tggccctctt aaggtgtcct ctgaagctct tcttagtgtg 159660
cccttacctt aaatattgtt ttcgaagact tcattttcag ctaacttcca ctttaaccct 159720
attcactctc cttagctgat ctcagctatt tcctgcatgt atcctaatgc acaaatctcc 159780
tgagctccac acatgcaggt tgaaggactt tgtgggcatc cttctagatc acataggccc 159840
ttcaaaaatg gcatctttgg gcagtgaggg gtggggaaag atggcatctt taaatctgaa 159900
tttatcacct aagcagtctc caagcctcat cattgcccta aatttcatca gtaggtacca 259960
caagcaccac ctaaacccaa aaactatatt ctttctttta tctttggtac atctaagaaa 160020
ccattatgtt ctattattcc acctcctaaa tatcactcca ctgctgctga tttggtgcag 160080
tcctccttat tcactgccct ctggccttct actatccttc aggaatcgcg aactctttgc 160140
agtttcccgg tgcactattt tccctcctaa ctctggacat ttgtttgcac atggtatttc 160200
tcccacttgg aatatcctcc aagcaggtta catccatggc agatagcgta tttctctata 160260
agcctgcagt tcctgaagcc tctctctcac ttccaggtta agtaggttgt tctgaacagt 160320
atttccttcc catcttttat agatccccat tagagtactt ttttcgtttt gtatggtaat 160380
tgtttggagc cacatggctg cctatgtttg catcccgcat gggtgccaat gaataaatga 160440
acaaacaaat atcgatgagt gaaaaagcgg aagcctccca aaagtagtat gttggattaa 160500
gactatccca tgatgtaaaa agaaaagaaa aaaaaaactt tttaaatcta gagttttaaa 160560
aaacataatt cctacatgga ttattttgag ccgattgcat ttgatttgga attccatatt 160620
taactcagag attatttttg ctaagtacag acatctttgt ttacctggcc agaaagaacc 160680
tactgcctac ttggtataat ttgaaaggtg ctgagggtcc ccaagccacc tctatccaaa 160740
actggcctag gtacctgtac tctgccaata gagttacccc tttaaatgtt ggcaggatgt 160800
ccggaaaaac ctgttcaaac ctctggcagg ggtaatgtgt acacatttcc atgtcctctc 160860
agatttatca aaaacaggta aatcttgatt agcaggagaa ggcgggtcag gcccacaaag 160920
gacccattct tcattttttt ctattttcag gattcattct ctgtgcttct cctatcttgt 160980
ggttctcttc cttagcaaaa agaggctaac tgagtcatag atgctgcttt tcatgctgaa 161040
cctctatcat ttatttccct tatatggctg agaaattcat gttgatgctt gcagtcaaaa 161100
atccttcact taatcttgaa tcatggctac ctgtgggtat ttgctgacta gccactgtgg 161160
gttaggctgt gggagaagga agaaaaggac cggaagtggt gctctgagaa tagtgatggg 161220
aatgctttca gaactatagt taggggtcaa attcatcaaa taagtaggtt tcaaagaaaa 161280
aaatgtgtag actcacttta gcccaagcta aataattctt aatccagaat ctaacttatc 161340
tggatgtttt gtctgctaat gtgtgtctct gtagtaatgg ttcagggagg gcattttggt 161400
ttcacattat tctattacta cttgcccttc ccccatttct ttgtggtttt ccctctcact 161460
cattccttct tcagttcttc caggtgctgt ttggatccat atacttatgg cagtggtctc 161520
atatatgtat gaactgatta ccaaataccg cctacttctt gccggagtca gtttttgctc 161580
ctgtaaaaac aaattataac cctacactca gcaacttcag agaaggtaac ttagcgcttt 161640
ggtctgtttc tctctacagg taaacaaaat aaaaactcct aaacaaatag atgagctgat 161700
tgaaattgag agtgacacag gaacatggta taggaggtgt tttgttcgga tccgcaccga 161760
gctgtacgga gtaagtaaca agtcccatga tgacctgcat gttaatttat tgcatttgag 161820
gttagtcaga agaggggttt actgggcctg ccctatgagc gaggttcttt cctgctgccc 161880
aggacacctg actcctggac tagagccagg taattggatc ccactcgacc agcatttagc 161940
caatacattc catctgtggg aagggaaagt gttaggccag gagtcttggc agtggcaggg 162000
tgccttcaaa tcttgagtta ccttgtccat tgacatccag gaagctgaag aatgaaaatt 162060
ccaattacct ttaactttta cattttaagg tgaaatgaat ccaacccaat gacaactcag 162120
cattttttat aatttggtgg aaggggtcgg ggagtggggt tgactaagta actgcatgag 162180
aaaactaact ctctaatatt tcacagattt ggttgacttt tatgagatgt ttcaactacc 162240
cagtcaggga taatacaata aagcttacaa ttgtttggtt caccctgtcc tttgggtaag 162300
48
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
tgatatttaa attcttggca agcaaaatcg ttcaatgtcc acattgtaac tcctagccat 162360
gctgctttct tttctgatac tgcttgatgc tgttaacctg ctaaggaaga aatttttttc 162420
agcttaggtt tgctgagagt tatggttttt ataaaccagt tattttattt gttttgtcat 162480
aaagctaagg tagcaaaacc aattgtatct gcaaaatcca gtaaggccct ggatgatttg 162540
CCCCatgaCt tC'tCtgtCCt gtCCtCCttt gcccagcccc agccatacca gcctctctgc 162600
tcttctttga acattctaga acagttccca tctcgggcct ttgcggtgtt cctctgcctg 162660
agttgctttc ccctccagac atctgcccag ttcatttctt taccctcttc aactctttcc 162720
tcagatgtca ccttctcagt gatgtcatcc tctcttttta aaaaattgca gcccctccac 162780
actccctatc cccttctcag ctctacttta ccccacagca ctttctacct tctaatatgc 162840
aaaacactcc tttattctgt ataaagagta aagctctggc ttccctctct ggaagctcca 162900
tgaggacagg ggtttttaac tgccttgttc aaagctgagt gtggtgccat ccacctatat 162960
tccaagctat tctggaagct gaggtgaagg attgcttgag cccaggagtt ggaagcttaa 163020
gagtgttata atcacacctg ttactagcca ctgccctcca gcctgggcaa catatcaaga 163080
ccccatttct aaaaaaataa taataatttt gtttatgtct tgttcactga tgtatattcc 163140
tggcatgtaa tagcagctca ataaatagtt gttgccaaat gaattaaact gccatatatg 163200
tgttccagga aaagaggaaa tagggagaaa ataacaagat tttatttgtt aatcgctgac 163260
atagtcactt tagaatgtgt gtgtacaact ctattgttgc ctaatttgct gtttagaaaa 163320
gaaaaattag ataatactct acactttcag gtcagcagga actcagtacc ttatagaacc 163380
taacatttat gttgcaagtt tcatctctta acatccctaa atcaaaccag gcaagtgatc 163440
taaggtgacc actctttctt agtgcctgag gaggtggaat acggagaaag aaggaagctg 163500
ggagtcagga gacctagcag cccattctgc tgggtgactt ggttagcttg tgacttggag 163560
acatttctca gttcctcatt tctaaattaa gagggttgac ttagaatatg taaagtaatt 163620
cccacccaag ctctagaaaa tccattaatc tctgaaggaa gggaaggcta aatattaacc 163680
tatattaagt gcttactctg tacctaagca aggggttaaa gacaaagaag acaaagataa 163740
gcaagaagga agtcctgctg tcaaggtgta aagagtaatc atcattatct ggcttgttta 163800
ttttgtttac taattcatta ttgatcatat tttaagcttt atgatttgtt ttactgctat 163860
atccctggaa cctcaatagt gcctagtatt tagtgcgtgt tcggtaaatg tttgctaaat 163920
aaatgacaaa ttagccaaaa gccctccaca tcattgatga tggatggaca ttccaaggta 163980
atgtgccctc ctcattgttg ccatttcaag gtttgtgccc catgtatgtg actagagaac 164040
aatgccataa atgttgtacc cctaagtcat ttcttcctca ataactaccc tcacaccccc 164100
acagacatta ctatgaagaa aaggggtggg aatcctgagg cacagatgtt tttggaatcc 164160
taatgacaat gatgataaaa acgactcata ctgtcttctg agcactttct atgtgctagg 164220
tacttgacat cccatttaaa ctgagcaacc acccaggaag gtggatattc ccatcattcc 164280
aatagtagag atgggcaact gaggcttaga taactggccc aagtttaggc agctagtaag 164340
cgacaagacc aggattcaaa cccagcctgt cctcctgcaa aggctatgcc catactctct 164400
tctttccctg tgtagctaag tgtcactggg aataaactag agcagtggtt ctcaaacttg 164460
aacgtgcatc agaagaatcc ctggaggact cccctgccaa agtttttaat tcatcaggtc 164520
tcactggaac ctgggtttgt gggggatgat gatcctgctg gtccggggac cttatcttat 164580
cttaggaggt actactttag agcagcacag tctaatagag gtatcgtgtg aaccacagat 164640
gtcatttcag attttctggt agccatacac acacacacaa agtaaaagga aatagataaa 164700
attaatttta atactacatt ttgtttcatt cattctctct ctcttttttt tttttttttt 164760
ttgatggagt ctcgctcttt tgccaggctg gagtgcattg gcacgatatc ggctcgctgc 164820
aacctccgac tccctggttc aagctattct tctgcctcag tttcctgagt agctgggact 164880
acaggcgtgc accaccatgc ccggctaatt tttgtatttt tagtagaggt agggtttcac 164940
catgttgacc aggctggtct cgatctcctg accttgtgat ctgcctgcct cggcctccca 165000
aagtgttggg attataggcg tgagccactg tcatttcaac atatgatcag tataaaaatt 165060
attagcaaca atttttacct tctttttttc cgtaagtctt caaaatcctg tggtatttta 165120
tatttatagt atatctcagt ttgaaccagc cacatctcaa ctgcctggta ggcacacgtg 165180
gttagtggct tctgtactga acagtacagc agcatgacac agctgttaga ataacacatt 165240
agaggtcaag aggcagaaag gggaaataac ttggaatctc tgagaataac acgaatgtcc 165300
tcttcccagc ctctaactac cttgggaact ggcattctac ccccaggata gcagtatcac 165360
agcagtgtag gaaccaagaa agattcccca ttagcaaaag aggtcattct ttaggggcta 165420
ctcaccgtgc ttggatcccc aggagtttgt accaccagac taggagagag agtttctcag 165480
agcagactgg aatgatgaga aagatattgg ggccagggat gtcagatgtc ttactccatt 165540
tctgctgcta taacaaaata ccacaacagg taatttgtaa ataataggaa ttttattttt 165600
cacagtcctg aagactggaa gtccaagctc aagctgctgg cagattcagt gtgcagtaat 165660
ggcacagtct ctgcttccaa gatggcagct tgtcgctgag tcctccagag gggacaaata 165720
ctgtgtcctc agtggtagaa gggatggaag ggaaaaagtg agagaggcag caccctcgca 165780
cctcttttta taaggacatt aatctcattc atgaggtctc tgccctcata acttaatcac 165840
ctcctaaagc ccccacctct aatattatta tactggggtt taagttccaa cacataagtt 165900
49
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
ttggaaggac acattcagac caggcaccat gtatgggcat aggatcggat gctttctgca 165960
tgtctcttcc caaaggaagg gtttaggaat tacatacctg gggttctcca gctaaccata 166020
ggccatctct caagcttgat ttttgcagat taatgctggg ggagagggac acatggattg 166080
ctgtatgatg aattactcat aatgttatag ggagccagcc attctccggg aactattacc 166140
agtctgaaaa catcagggag ttgctcattg cctctggcat gctagtgcct ttacaaactc 166200
acatttcttt gtctttgcag gtactatgga ttatccgttt ggttccctga tgtcattaaa 166260
cctctgcagt ccgatgaata tgcattgcta accagaaatg tggagagaga taaatatgca 166320
aatttcacta ttaactttac aatggaaaat cagattcata ctggaatgga atacgacaat 166380
ggcaggtcta gaaacttgaa ataatttaat ttgctaccta ttcacaaaaa cttatttctt 166440
cttagctttc cctgcactga aacaggcata gttttttgtt tgtttgtttt agtttgtaca 166500
tttagtcatt ttcatgatga atttcaatac ttcttaatag gctttaaata aaaggtacaa 166560
atgtatccat tacttcactt aaaactattt ttgcagctaa gttccttcat atttattagg 166620
tagtggactt gactggtgtg gaaggaaaat ttagagcaat gttggagagc aacaaaaata 166680
atggaaggta agagaagagt gaagagtatg ggaaaataag acaggctata ggattattta 166740
attcacagaa gaaatctgaa atgtgattat atgagtcttc tattcctgag aaacagagcc 166800
acttttcaga attcatcctt cccaatacgc tgaatccaga attgaggtta aattgcatca 166860
ggaagagttt caattagaga acaggaatca ttccccaaag taaagggctt tgaaagacgg 166920
aagggtctcc tttatgggat ctgaaatact tcttgatgac aagagagaac tgactcaggg 166980
aggtggtcct gggtcattct tttagagatg gaaaagagta gaccaagtac ttgtttatat 167040
ccaagtattt ttgttgcctg aacaaataat tgccagtgta agttcatgat agtttacatc 167100
attcttagga agttgtccca atttcaaaag tgataaaata aaactctaaa ataatcaaag 167160
ttccccaggg caaaaaccag ttcagaaaga tgaaaaagaa ttcagtggga tgcttatgat 167220
ctatgtaatg gaaaagaggg ttactaacta ggaggtaatt gttcaaatta gcaattcaat 167280
gaaggaacta atcatcatcc aagccaagtg ggatcacctt cactctccca cataaaatac 167340
tgacttgaat ctgcacttag catttcacag aaagcttagt gtcaagtcct ctgtgatgcc 167400
agggtgaaaa ctgactttca aaagtattgt tactccatta gaaatttcta atatatgttt 167460
gtgtctctga cagtcatatt tttagtaata tatgttaaag ttctaaactg gatactcagt 167520
cactgcttag aactttttta cattagcaat ataatgatta ctaattttaa tagtattaca 167580
gtttttgtaa aacatatgtt acacatttat tttaaaagat gatagaaaca atttttttgt 167640
ccttcaggaa aatttgaaat accattaacg aaatttttta aaaaatagct tttttattca 167700
gcatcataaa atggcaactc tttatatact gatacggaat aacctccaag gtagactaga 167760
aggagataaa gcaggtgtac tttagacaca gaatgctcct ggaggagcac acaagaaact 167820
gacaacaatg gttattctgg gcagaggaac ttgtggcctg gggatggatg ggaaaagact 167880
gactttttcc attttcaacc tttcgaattg tgcaccatgt gtactctctg aattatccat 167940
tcaaaaaata aaactgattc attccctttt ccaaattcag ttccatgaca gaaaaaattg 168000
ataacttttt aaagatgtgc cattatttag tctcttttgg tgcatttaga tgtttttact 168060
ctgggtccct ggagtgggga ttgaaacgag atcattatta ttatttccag gtaatgtacc 168120
atgggaatga tttctttttt aagctgattg gtctgtgagg ccatgtgcat tttgaaccaa 168180
cacacgtaat tgggtattag ggagagaata aatttgaaag tactcaatag ttagtaaact 168240
gcttttaaat caacctactt acctagaagg atcagtccct ttctgcccct aataagagtg 168300
gtattgacat aacaactcct tcccgaggag aacaagcatt ttcctctcag tgactggcta 168360
agtgtacatt catagtctct taataaagta ccaggacacg ttaatgaata tttatctgtg 168420
tctgctggga gtccttatca atgctgataa ggcctggatc atggactctt ctaagatgtt 168480
attatcatta tgactgttac catttttacc acgtagaatt cccccttata atacccaaaa 168540
atatacctga acaggtttca tcattcttca tgtttccaaa aataagcaga gttctcataa 168600
ttccttttgg ttcaaggact atctgttggc tgtgagaaga aaaattatcc tggaaatgag 168660
cgtcacctat gagtttcttc aaccccaagt cagttattcc tcacataaat gaaacagaat 168720
tctcatgctc tctcttcagg ataattacag gtccctacac tagaatgaag aagacttgga 168780
ttttcagcta gggaaaaata aagttattaa aagatcgggt aggaggacat ccttaagtta 268840
gagccattaa ccccaaatag agaatattca aataaagata tacatgaaga gacgatagga 168900
ttaaaaccta aagaatagcc ccaggtaatc cccaaactat cttcagttgc ataccccaca 168960
aaacttttta ccaacattac aaaaagctaa gggaagttag gagtgtcatc tcttttcctg 169020
gtgaaaaata atattggtaa gcaataagat agccaaaggt cagcctggat gagaaaaagg 169080
ggaaaaagca gagagctaag aaaatcaaga actctgtgtt ctttataatt.aagacagtcc 169140
atagtggggc attccatctc taaaactatt tgacttagct ttgaactttg atggacacag 169200
tcattgattg atatgaattt ttctgtgtgt tgggcagatt cataggggtc aagttcaaat 169260
ctgtaacttt caaagactct gtttttaagt cctgcacctt tgaggatgta acttcagtga 169320
acacctactt caagaactgc acatttattg acactgtttt taacaacaca ggtaggtgtg 169380
ctacttagta ctgcctctac ctgaggcctc tccaaagggc ccactgtgat aaccatgtga 169440
taatgtgggc agaggcttat gcgggaagtg gatattaaag aataaatgga acaagttctc 169500
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
taaatcaggt tggatcaaag gttggcttgt attgtttgtc tttgtcagat ttagtgcaaa 169560
tggagcttgc caaagttcaa aagttggcaa caacaacaac aaaaaaatcc tttgatgaat 169620
aaatttttaa agcaaagacc taacctgtag tttatttgtt ttgataatcc tgggttcaaa 169680
atctggtgtt tgtttaaatg cctcaactct atagccaaca gagatagaca acatgtggct 169740
atgctcagat acatatatat tttggccaaa tccatataat ggggtatttt aaactcccca 169800
gggggaaaaa aaaggaaggc aaacaagcat tgattgaaca cctatatact gttttctact 169860
ctaggtcatt agacattcat tatctcattt agttttcaca gtagctgagg cttaggaaag 169920
ttaagtaatt tgtccacaat tatagaacta gtaataacag atgtttgaac ccagatttat 169980
ttgatttcaa agcctgtgcc ttccactttc ccactcccaa acattggtct gtgtgtccta 170040
taaaatgaat ccagcataac agcgtggcat tggcaggagc atcagcattt gccctgttgc 170100
cagacctgcc ttcttagtgc tcagacaagg gctctgccta gctccaaaga agaatgaggt 170160
caggtgtctt tctgtttttt gttttttgag caatgctatc tacctgtctt ctaacccatt 170220
aaacctcaga agacgaacat tttgtcagat cttgatcagc aaggaagata gtttcaattc 170280
tcacagcctt ttagcagaaa ccaccaccat ctgcaactct tcttggatgg tttcaaattc 170340
aagaaaagat tagcaatgaa tgctggtgac cttcagatga ggctgactct tgttcctcag 170400
tagattcatg cacatagact ttaaattata aattcagcct caggaaagac ttgttccttc 170460
tctgtgccct ctagtctcat aaaatggcaa tgataaggct gactcttgca cttttttttc 170520
cttagagaca gggtcttgct ctgtcatcca ggctggagta cagtggcgca atcacagctc 170580
actgcggcct caaataaaaa attattatta ttattattat tattattatt attattatta 170640
ttatttttgt agagatggtg tctcactatg ttgtccaagc tagtcttgaa ctcctggctc 170700
aagcaatcct cctgcctcat cctcccaaag cactgggatt acaggcatga ggcaccacac 170760
ccagcccatg cacattttga aacaaattgt ttttataatt gtagtatgga cactattagt 170820
actataatga tagtctttat gtaacatgat aattaggaca tagtaatatt gtaaaggagt 170880
aagcaagtaa tataattagc aatggccgac cataggaaat cagaagataa ctggacccct 170940
taatttaatg tattccttta ttcagcaaag gtatctcatc attcacctac tttatgatgt 171000
gaagacccag actagatcct atgctttggc cagaagaccc aaaaagtcaa gctagcatag 171060
gcctgaaagc cctcaaggaa tttacaatac tggtttcctc tccgaatcct ccataggctt 171120
tcttatactg gtgcatgcct ggtaagagct tgatgaattg tctttgttgt tgtttctaca 171180
gattttgagc catataaatt cattgacagt gaatttaaaa actgctcgtt ttttcacaac 171240
aagacgggat gtcagattac ctttgatgat gactatagtg cctactggat ttattttgtc 171300
aactttctgg ggacattggc agtattgcca gggaacattg tgtctgctct gctgatggac 171360
agaattgggc gcttaacaat gctaggtatg tactcagttt catgtcaaat aaaagagctg 171420
cccctgcaat ccagagggac cctgtttccc cctgtactcc catctattag ggatttgcat 171480
ccaataagga gaaatccttt agaccttacg ttgggactaa attatagtga aaattatgtc 171540
taattacatt tcatttgagt ggtttccatg agttaacaga ctgaaaaact atgtgtaaaa 171600
tgtatatttt cccactcaag tagctgagta aaagatgata tgcatttcat actttctgtg 171660
ttcagaggca gttagagcct ttcattcagg tgaatgcacc agttcttgag ctttatggag 171720
cccatcctgc agcttccact tagaattcag ttttcttgag caatcccaag gacccaaggg 171780
ttcttgcttt actaagatga caggaaacat tccagccttt tgtctgcatt gttggcaggt 171840
ggctctatgg tgctttcggg gatcagctgt ttcttccttt ggttcggcac cagtgaatcc 171900
atgatgatag gcatgctgtg tctgtacaat ggattgacca tctcagcctg gaactctctt 171960
gacgtggtca ctgtggaact gtaccccaca gaccggaggt atgttgaaat gggcctctag 172020
taagaggcgc tctaagccct gtgagaccac tgaaaaatta ttttaaaaca acccaaactg 172080
gctgggcacg gtagcttatg cctgtaatcc cagtactttg ggaggccaag gtgggtggat 172140
cacctgtggt caggagttcg agaccagcct ggccaatatg gtgaaacctc gtctctactt 172200
aaaatacaaa aattagccgg gtgtggtgcc aggtgcctgt aatcccagtt actcaggagg 172260
ctgaggcagg agaatcactt gaacccagga ggcagaggtt gtggtgagcc aagatcacac 172320
cactgcactc cagcctgggc aacagagtga gacaccatct caaaaaaaaa aaaaaaaaaa 172380
aaaagccaaa ctgtcacttc cccaggaaag taaaaatcca aaaatatctg gcttcacaaa'172440
gttgtaggtt ccactcatat cttttaaaaa gaaaatatag gggaacatat gtgctagaag 172500
gaattaatag ggcaatgaag ccagtgaagg aatttaagtg tgtccttaac ttagtttgta 172560
ggctaacaat gtgaggaagc tgtaatgaag cagtgaggat tagagtttaa tagcctatct 172620
tggcccacat tttaagtggc taaccagtcc ttagagggtc gtttccttag gtttgcagat 172680
gtgtctgcat gagctctgca ttaagcaacc acaaagaggt atctagaaca.tatctcttga 172740
ccttcctggt ttgttggagg aaagccaaac acatttctag ttataaaaaa caattgtgct 172800
gggcctagtg ggacatgcct gtaatcccag caatttggga ggccaaggca ggcagatcac 172860
ctgaggtcag gagtttgaga ccagcctggc caacatggtg aaaccccgtc tctactaaaa 172920
atacaaaaaa aaacctagcc aggcatggcg gcgggtgcct gtaatcccag ctactcagga 172980
ggctaaggca cgagaatggc ttgaacctgg gaggcggagg ttgcagttag ccaagatcat 173040
gccactgcac tccagtctgg acaacaagag tgagactcca tctcaaaaaa aaaaaaaaaa 173100
51
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
aaattgctcc caatatggaa acacagagac tcatatcaga gaaagggcta gggattttgg 173160
gtatagcttc agcagcttga atagctaaat taaaataaaa gaagctctgc tttaccattt 173220
gatatgagct aagacaaatg gactttgcct tcatgtacca ctcctgtgct atgttgccat 173280
agaagtcctc tgtgccagag ggtggagtca gtattatctg gactttgaaa tgtaggagag 173340
ttttttccta gaactatgcc aaaacatttg ggatatcaat ttgcaactga tcttgagtgc 173400
ttcctctggg ctcattccct ttttttattg accccaaaga ggtgtattct gtgtgtgtcc 173460
tgtcctgtct ctaacttata tttcttcttc aacaacatct attactgagc ttgcagaacc 173520
ctgtaggtac ccctccatcc atctccctgc agaacatcta acactcaacc ctggatgtct 173580
gcttatcaaa tattctagtt cagaccctca gtgacccacc tattaaatat ttaggctctg 173640
aaaggaacca tacaatcttt gtacggcatc tccttcattc attcaacaaa tgttaattct 173700
ctgtctttgt gaacgaggtc ctgccagagt tggtgcggcc aaaagcaaga ttaagactct 173760
gcagcttcat tgagctttct ttcatcaacc tctcattctg caaacagagc ttagatgtgg 173820
attacactat acgtttaaaa tgaacccctc aattgcttga gcccattgtg ccactgcact 173880
ccagcctgag cagcagagag aacctgtctc aaaaaataaa taataaaata aaataacctc 173940
ctcaatgtta aagcaaggct aggtcatagc ctccttcctt tttcttgtat actataagag 174000
tgtgatactg ttttaggaga atcagctgct tatccctctg ccataaggaa tagaaggatg 174060
agtattcacc taggacaaaa atctggctta aaaaaaaaag accctcataa attgaaattg 174120
gtcaggtcct tcctgtgaga agtacagacc tacggtgttt ctgttgtcag aaaggttaaa 174180
tatcaattga aataaaaaca aatgttgctt atttgtatta attcaggtcc tccatgaagc 174240
agatgtcagg atgggattaa acatgcaaaa atgttattag aggaaatgac tgtgagagaa 174300
gatggagagg gagtgaagta tgggagagct gcaatgcaac tctaagcctg agcgaaggag 174360
ggaggaagga gtatttgggt ggaatctccc agactatcca gtaatctaag gaaggtttgg 174420
caaggctgtt ggagacccag ggatcatgac aacatcagcc tcagaggagt cctgtgtgac 174480
tcaggaagca gcctgcagta atagtcctgt caggtccagt cattgtctgg gagcagcatg 174540
tgggaaaaca cctctgtgca caccagcacg ataacacaaa ggtctgccag gtgcattctc 174600
accgcggtca cgctgttgct ggcagacgct ctccagcact taagtagatg tcatcagata 174660
atcacaggaa gtcatttcct ttggtgatct agaagatgac ttgtgctcca gagcagcgtg 174720
accatggcca tggtaaattc gctagtatga gctaattagt gagtatctga aaatgtttca 174780
ggctaaaacc tcttccacca tttccatgtt tctctgggtt gcattaggag gtctccactg 174840
atctctaact cagctattag aggaaaatcc caaccagttg agacactcta tagcttcatc 174900
tccccatgga ggcaggatag aatacaagaa tacaggaata gaacacaaaa attctggcta 174960
gaggctttac ttcattctct tgtgttgctc taacaaagta cctgagacta agtaatttat 175020
aaagaagaga aattcatttt ctcacagttc tggaggctgg caagttcaag atcaaggcag 175080
gtttggtgtc ttgggtgaga gctactgtct gcttccagat gttgctgagt ccttcagagg 175140
agacgaacac tgtgtcctca catggtggaa ggcagaaggg caggaagggc caaacactgt 175200
gtgaagcctc ttttatgagg gccttaatcc cattcaccag ggagaagccc tcatgaccta 175260
atcatctcct aaaggccccc ctctcttaat actgggattt gtattggttt gttcttgcat 175320
tcctataaag aaatacctga gactgggtaa tttataagaa aagaggttta attggctcat 175380
ggttctgcag actatacagg aagcatagga agcatagtgg catctgcttc tggggaggcc 175440
tcaggaagct tccaatcatg gcagaaggta aagggggagc aggcacatca catgtaaaga 175500
gcaggagcaa gagagctcaa ggggaggtgc cacacacttt caaacaacca tagctcacag 175560
gaactcactc accattgcga ggacagtacc aagggcacag tgctaaacca ttcatgagaa 175620
atctgccccc ataattcaat catctcacac caggccccat ctctgacact ggggattaca 175680
ttctaacatg agatttgggt ggggcacaca tccaatctct attaggatta catttgcatt 175740
tactgtgaaa acactaatga ggacgttgtt cattatagga cacatcttct tctactcaga 175800
gtctaccatg aagactgaac atctcagggg tgggttctag gaatcatttt gtattcctcc 175860
acccatgctt tgagaagctg ttatgtgtcc tcacttcata tgataaaaat tgtgtaattc 175920
ttcacatttc cttctctgtt gtgcaatcag gaagcagaga gcctgtgtct ctgctaaatc 175980
tgcatagtcc agatacttgt ttcactcttc cttatcttga tttgccccct cttatgccca 176040
ttttattaat tttgtgtctt ttataagtta tcctacctag atcgtttgat aaagaggcag 176100
gaaattttgg aattaaatct agatgtactt actgaatagc acttttctgt gccaaaaaaa 176160
atgaaacaac catttttcca ttctgtaaat gccatcaaca tcctcctttc ttcaaaacct 176220
cagtatcaaa aatgagcata acaattgcag tcttaaacag atgcatggtg acattctagc 176280
tttttcccca gattagagag cacttacaca ctgactggga catgatgatt taagggacac 176340
aaaaagaggt gggaggggac tgaacattcc aaccctctgg acatatggtt ggtttttctg 176400
gagaccagcc caatccagaa gctatctagg atcccctcag ccaagagtca tctcattagc 176460
ctacaaaaga cactcttttg gtggagcagg gggaaggatg gggtctccct ctgttgccca 176520
gagctacaat ctgtaaccag agctggaatg cagtgattca atcatagctc actacagcct 176580
caaactcctg cgctcaagtg atcctccccc atcagcttac aaagtagctg gaactacaga 176640
tgtgtatcac catgccaggc taatttattt tatttttttt tattttaaga gatggggtct 176700
52
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
cactgtgttg cccaggctgg tctcaaactc ctggcctcaa gcaattcttc tgccccagcc 176760
tctgaagtag ctggtattat aggtatgacc cattgtgcca ggctccaaaa gatactctta 176820
tccaagggtt ttaggagctc aatgccggga actggggatg aagaccaaat acttattctt 176880
tacttatacc ttatatttaa ggctggagat tattactaaa gtgcaattta ttatccagag 176940
gatttagcta tgcaaaaggt gggaaactgt acccctggac agcaaatcaa aaacaaatct 177000
acattcccta aaaccttagc tgctagttat aagcatgacc tcaaaaccag aaaccatcag 177060
ggagaaggtt aaaagaattc tctgggtaaa acaaaatgtg aattccacac agcaaaaaat 177120
accataggca catataaatg ataaattgag ggcaggaggg ggttgaaaca gaaatgaatt 177180
aataacctga atatataaat tggaaattca taaagaaatg tcaacaaaca cttagcatat 177240
aaaaatgctc agcttcacta atagtaaaaa ttaattacaa attaaagcaa ttatgacatt 177300
tagctttctc ctattattca gagaagacaa tatttaaaag atagataaga tccagtgttc 177360
tctggggcaa gtgaggggtt actgctgagt ctgttgtggg aattctgaaa acagatacca 177420
catttgtaga gggggatgat ctacagtctt aagtattttt tcccaatagg aataaactat 177480
tgattcctct ggttgctttt caggtttttt ctttgcctgc catggatttc tttgggttta 177540
ctatgtttgg ggtttgctca gcttcttgaa tctgtaggtt tacagctttt gccaaattta 177600
tgaggttttc agccattatt tattttgtca aaaaacaaaa ttacaacaaa tttagtgtaa 177660
agagctagtt gacttttatc tgctattcta gaagcaggca acacctcact ctataaaatg 177720
gggtgagtgc tctaatgagc tgagcagaga aagttggctt cataggcaga aaagctctaa 177780
agagagcaga tacagagaac aaaaagcaga tagccctttc aaagtccctt tctgtaaagg 177840
gctaaaacag aggggacttc cttattatgc tcatttgggt tggccggaat cacctgtttt 177900
ttggaaaact ggctcttttc aaagttcaat ttgattaggt ggcacttagc ctgagtgact 177960
ccattctgct taggactggt ctgctgtggc ctaggtgcag gagactagcc caaaacaatg 178020
gcctcccgta aaatttaact atttgaatac tttttcagtg ctgccctttt tctcttctgc 178080
ctgaactcca gcaacataaa tattagatct tttgttatag ttccgcaggt ccctgattta 178140
gtttgttttc tctgttcttc actttagata gtttccattg ttctgtcttc cagttcactg 178200
gttctttcct ctgtctcctc cattctgctg ttgagcccaa ccactaagct ttttattttg 178260
gttatcatat ttttcagttc taaaattttc atttagttct tctttatttc ttctacttct 178320
ttgctagggc tttctatttt ttccattttt gccgtgatgc tttctaaatt tcatgtgttt 178380
caagtgtgtt catagttgct cattgaagcc ttattatggc tgctttaaaa tattcatcag 178440
ataattctaa catctctgtc atatcagtgt tgccatctat tgattgtctt ttttcattct 178500
gtttgaaatc ttcctgttta tttgtatgac aaacagtttc tgattggaag cagtcattta 178560
catattatgt tatgagaatt tggatcttat ttaagccttt tgcttgaatt ggctttctgt 178620
gactcctctc tggcaagcga aagggaggca ttgccttatt attaccaggt ggacgtagaa 178680
gtccaggtac tccacttaag ctccagtgac acccaagagt gggcattctc attactgctg 178740
ggagtccaca ctcccttcct gatcatggat accatgggag ggagggaagg aggagctcat 178800
taccacccag cagggatgaa agtcccagct ccttacttac ccttctctga tggcatccca 178860
atggtagtgt tgacaggtct tcttagtttc acaagggtgg aagtctaggc taccaccaga 178920
cctttgctga tgtagctggc agttgagcta caattttttc ctggtgtttg gtctgaggag 178980
agaggttatt atctaaaagt tttctatctt gctagactgc cgttttcctg gtccttgggc 179040
tagagagagc aggctttggt tgtggctttt ctttttctgt gcctattggc atttctgggt 179100
tggcaacttg ttcgattcca agtctgagat ctatggagca gaaggcaaac gcagggaact 179160
ccccatcccc aagctcccta gcagttctgc ctttttcctc tttcagtctt gttctgtttg 179220
ttttatatat aatgtgcagg atattttgtt gtacttatca agaagaataa gaaaagtata 179280
tctattccat ctcagtgtga attttaatac tgtttgttta tttactcaac aactattttt 179340
ctgagcacct atgatacaac cgtgagtaaa tcagttgtgg tctttattct cataagcaca 179400
agcaaacttc agttgtaata tagtggtcag ggttattaca tgggaagtgc atgatgctca 179460
gagcatcaga gagggtttct tgaaagaagt accatttaat tgagatctag taggtgataa 179520
ccagtcaaaa agatggaaag aacatttcag ttagagcaaa taccagctaa caagctctgg 179580
aagtaaaaga gttactcaaa gaacaagtca tgaagactct ttcacattgc cttattgtgt 179640
ggggacgggg agactttgag ctgtcttaca gagaatgaca agatcattct tggcctttag 179700
agagctccct ctaatggcca gaaagaaggc agggaggctg gttaagacac ttaagttgta 179760
ggttggaggc cgggcacggt ggctcacgtc tgtaatccca gcactttggg aggccaaggc 179820
aggcggatta cgaggtcagg agatcgagac catcctggct aacaaggtga aaccccgtct 179880
ctactaaaaa tacaaaaaat tagccaggca tcgtggctgg cgcctgtagt cccagccact 179940
cgggaggctg aggcaggaga atggcgtgaa cccgggaggc ggagcttgca gtgagccgag 180000
atcgcaccac tgtgctccag cctgggcgac agagcgaaac tccatctcaa aaaaaaaaaa 180060
aaaaaaaaaa aaaacagaaa gaaagaaaga aaaatgtagg ttggaaaggg gaaaagtcaa 180120
cactcaacat gttcacagat atttagggat acattcaaca aagcttggtg cttgattaga 180180
tgttaggggg aagaaaagag gttgtcaaag aaagatgatt ccagtttctg cgtgtacagt 180240
tgagtgaatg gagatggcag gttttgttta tgttttttaa ttttaaaaac cacctgcagg 180300
53
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
agcacaatgg cattttgagg ctgggacaca ggagggacag tgggtctggg ctgtggtcaa 180360
gaaggtaata agttccattt tggacatgtt gagtttttac atcaacctgg ggctgtccga 180420
ttggcatttg aatatttttg tttggatgcc agaaagatac ctgagctggg gatagagagg 180480
tcattcacta tttcattcaa aaaaaaaaaa tgattatgga atgcatatcc tgtgccagga 180540
actattttag tcattgggag acagcagtga acaaaacaga cagaaagctt tgccttgata 180600
gagcttatat tctataatag tttgaggaag acagtccaac aaacagataa gtaaagtaga 180660
tgtagtatat tggatggtgc taagtgctgt agagaaaaat aaagctacgt ggagcataga 180720
gaatgctggg gtaaggagag ttacaactgt aaatagaatg gtctaggggg atgtgactgt 180780
aaagctgaca tttgagcaaa gacccaggca gacatctgga gagagtgtat tcacagcaga 180840
ggcaacagca ggtgcaaagg tcctgaggtg ggagtgtgag agaccagttg gaagagaggc 180900
aaagcttgtc tggctggagc agaatgacca aggtagaggg taagaagcca tggggtcaga 180960
gtggacaata aaagggcttt ggtcttcttg ctgagtaaaa tgaaatccaa tgtggggttt 181020
gggggcagat gagtgacatg aactgactta tgaagtccac agcaagaaat gtggcaagag 181080
gtaatggtgc caatggaggt ggtgagaaag tggttagatt ctgaaaatat gtctaagaga 181140
gagacctgga atagttcaag tcaaaggaaa cagaaaacaa aggcctccat agaaataaca 181200
tgtctgagag gaaagagttg gagtctgggg gtaaatccat caacgtaaga aaatccagag 181260
gtaacacaga tctggactaa gaaataacag acaccagagt taggatacca ggccaggaga 181320
caagagttca gtgagctaga aactcaggct ggcagagtgg aatacaaaaa aactagcctg 181380
gaagcaagtg gtcaaagtat taagatgtct tgatcttcaa aaatggtgta tgacattgag 181440
agaaggcttt gcaagttaga cttccctata acctctgtgc ttaggtcagt agggcagtag 181500
tagtttgcag ctggttctca atgacattca ttggactgac ttgcccaggg ctagaactca 181560
cctcaggtag aggcaagata tcagaactgt ctgcagtgga agttgaaggc acctctctgc 181620
tcatgcactc accttggtca gtcacctcta tcttgatcct aaaccttctt cccaactctg 181680
tagtaatagc ctgagattct tgccaaggcc agcttcccag tgctgtggta cgtctgccac 181740
aatgattgag atgcagtggt cgatgcagtg cccttctccc caaggctctt aattcttagg 181800
cgtagacttg aagaagccag gaatacttgt acttctgcag ggatgagtga ggtgaagcgg 181860
gaagacggaa aagcaaatgt tgcattcagg aatttgtcct tgattgtgag gctggaaaga 181920
caagattgct ttgagaagca ttatgaggaa agaaatgaag agcagggcaa aatgatagac 181980
ctttgaagtt tgagccatta ttgaatatac aagtcattgt atagtctttt aacagtttct 182040
acatcagccc ctcacctgtg tcaaagataa gcagagagcc tattatatgt aatgtagatc 182100
acaaacaatt gatgtggcat gagattcttc taatagacat gaaatatgaa tgtgacatga 182160
gccagacata gaatggattt tccagcttgc cgtaaatctg aaaggagccc caaccatcta 182220
acatactggt gtgggggagc ttccttttcc ctcatttctc cgtccccagt ccctgccacc 182280
agcaaaatta ctttcaaccc tctttgcggt agaactgaga acaccaacag aagatgcatt 182340
tcattttttg ggtttagtct gtgctggtca gcttccctca ataccctaac tgggaaaaat 182400
aagtcaattt tccctcatct cttgaagtct tcaaaatgaa agtatatgta atgacaggtt 182460
gaaatggatt ttttccatcc atcagggtcc ctacttttct ctccctgctt tgtgaaagtt 182520
gtaaatctct caaaagtcat aaatgtagct cccgcggtgt aggcctgttc cagctcattc 182580
cagttctagg ttttggcctt tatgtttctc tgcgtccagg tgttaacttt ctgctgccac 182640
tatgtggctg tggccaagtc ccctgctcac atttcctgtg tgcttatggc tcaggggcca 182700
cttttttttt ttgggggggg ggacagggtc tcactctgtc acccaggctg gagtgcagtg 182760
acgcaatctc cactcactgc aatctctgct tcccagattc aagcaattct cctgcctcgg 182820
cctcctgagt aactgggatt ataggtgcct tccaccactc ccagctaatt tttgtatttt 182880
taatagagac ggggtttcac cgtgttggcc aggctggtct cgaactcctg acctcaagtg 182940
atctgcctgc ctcggcctcc caacgtgctg ggattacaga catgagccag cgtgcccagc 183000
cattaggggc catctttacc cctcctccgt ggcgctcggg gccaggagcg cacaccttgt 183060
gagatgcagc ccacgctctc agggccgtcc tgcaactctg cacagactcc cagggccagg 183120
caagcctgtc tcatggtgtc gtttggccgc atgttcctgc ctttctctgc ctaccttctt 183180
cccttttctt tcccccttat gcccccttat gcatgcacag tttcagacat gaggtcagaa 183240
aggatttgag gatctgattg ttcacacctg ccgatccctg agagtcctgg gcagctgctg 183300
ccatcaccca gtgcagtggg attggtctct tccttgatga gctaaaatca aaacattttc 183360
ccagaattaa gcaacccaag cccttccctt aggactgtcc ttgttttgac agagttctct 183420
tgtttcctcc ccattcttca gttccgaagg tctgtcatct tggatctctt actgaccagc 183480
tgtgcgacct cagggacacc actcagccta actgggcctg gattgtttga gatttaaatg 183540
aggagtatga aataaatagc atctcagggc ctttctagct ctagaattct cttcactgtc 183600
gtcatctggt attctgcaat ggcaatttca ttcattcttt cttgaataca ttctacgtct 183660
aatataaaga taaatatctc acttatttgt gaagattttg gtacagagga atctttcagc 183720
tatcgcgact aaccaaccta tgtaatccac tccatccctg gccctctgct ctgcttgatt 183780
tttccttata tgccttatca ttatattata tatgtttatg ctttggctat ccagcaaaag 183840
tcagattcat tttgtctctg gtagttagaa gtagagggtg aagggggtgg ggaatatatg 183900
54
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
cttcctgctc ccacaacctt gccatcaggc aatatattac aatggaagca aatcctgcca 183960
agtgtactag cacaaaaaca gtcacaaagt ttgtttttga gctccaagga.agtccatctc 184020
gctgtccttg ggtactgagc aagcccttgc atgaaagggc catgtgcttg atatgctgac 184080
agcagggcca ccagacttgt gggccagcta aacaagggaa tgccccttta attgtggggt 184140
gcaggataag ggggctctag aatctggagt ctagaccagg agtaccattt caccaacttg 184200
aggctgcagg gcttggagga cgcttgtatg ttcattaggc taagggcaga acattctttc 184260
caatcattgc ttatacagaa cagcactcac atcatcagtg aaaaaatcac ctccgtggtc 184320
attaacaaac cttctcatgc agatttctaa aatgatcttt aaagcactaa gtaaccatct 184380
ctggcctgtt gattcttgat taatgtgctt taaattatat gggtatgcat tagagacaat 184440
ttatctatca atcatattgt ttatactgaa gtacttaaag gaaattatca cccagacctt 184500
atgatgagag gatgttaggt tgcatgtgta cttgtttaac tcactagaat gttagctcct 184560
tgagggcaag acattgtctt ccttatttac tgccataact ctaggtgaat gaatgaatga 184620
atgaatgaaa tatgcatttg agaattaatt tggctatgga tgatagtttt cctccccgta 184680
tggggtccag cctcccagct ttccagagtg agcatttggg ccagtgtaag ggagggagca 184740
ctggtcttca gtcctctctg ctgtcctttg ggtccctttc cacttcttcc atcacagcct 184800
gagacctcct gatccttcag ctgtcagctc cccgttcctc ataaagcatt gcctattaga 184860
gatccacctc cccctctccc tctacttcat caacaaagtg gaggaaagca ctgatctagg 184920
ggtcactgaa ttcaggtgca gtcactgcca gaaaaatagc tggataattt ggaaaaggcc 184980
tttcaccctt ctgagccttg gtttcctcat ctgtgtggtg gagggtgggt cgggcatgat 185040
gctggaaatt ttaaatgatc tcaacttatt taattctcat cccaatcctg taagagtagg 185100
tattcttacc ccatgttcag gtgaggaaaa tgcggcacag agaggggaag taacttgtcc 185160
tggatcactt agccttaaac ccagatgtct gactcagtgg tgctgctccc agcacagaca 185220
tatctacctc ccctcccagt aggtgtggga atctattttt aaacagctca cttggctgtt 185280
gggatgcaca gccatgttca tgaccttggc cacagatgaa ctccaagggg tgctcagtca 185340
ttccatcatt atttaccctg gcccaggccc tcactcccta gctaggttaa tcagataaac 185400
aagacataaa ggctcccgtt attaccttcc cagaatattt gatttcaaca ggaataagca 185460
gtcaaaagcc ccttatggtg gtggagatgg gagggagtta aagggctact tgggatccag 185520
ggattcctgt cagctcacaa cttgtaggtg agtccttctc cacttgagtg taaggaaggc 185580
cctgctcgaa ggggtgcagg gtgccatctt gccctcatta aggaacatgg ccaggcgcac 185640
acacacacac acacacacac acacacataa tagatactag ggtaagacag atttttatct 185700
ttctttgaag attagaaaac aattgcattc agtcttttct gacttaatat gaaattcatt 285760
ttttctctct gaaaaaaaaa atatgatagt ctggtatcaa aaaaacaaat atgacccaac 185820
taatttccat taaaatgtct attacaaaaa tcgacttcca acatatttgg aagatctcag 185880
gaaatctgtt agctcagtct ttcatttaat aagtaacata ggtatctgta ccggtcctga 185940
gaagaggtgg tgctgagtga ctacaaaggc tacccagctc cactccccca gaggtgacct 186000
ggacctaaac aaggctaatt ctggacaagg agatgtgcaa gattggtata aaccagccct 186060
tggaggggag tgggaagggt agcgtggagg gaatcaagtt gaggcaactg agtaacagtt 186120
ggaagagaag gaaacaattc aggttaggtg agaagctgcc tctggtgaaa tgaattaagc 186180
tctgtgctgt gagaagcaga ataaacaagg gagtttaata agagcctaag caggaaggga 186240
gaagaaaaaa gggatgggaa gcctttgggt tggatccctc tctctctcct ctccagtgtg 186300
caaagcatac tttgctggcc tcagtatccc ctgcatcaca tcataaacac tttcccttgg 186360
atgttttatg ctgcttgata ttccctaatg aacacacaca aacgaagcag gtcatatatg 186420
tgaaggtgtt aattgcagaa tcacttgtta ccccaaattt ggaaagctcc cattagccaa 186480
tggcatggtt acaagaataa tggttccatg ctaaataaat catttcattt taaacaaaac 186540
atcaacatga gaaaacagta atatgaatga aaaaaaagca gaaagcaaaa ttgtctaaat 186600
cctctgattt cctgtgattt atatgactgg aagtacacta aagggaattc agaaatgaaa 186660
accatttgtc agaaaggtgg agtggatgtt ttgcttttat aatttccttt gtatatgaaa 186720
cccacatcaa ttcaaagaat gcagacatcc ctgggaagac agcaggattc tcaaccctgg 186780
atacaccttc atgtcacgtg gagcttctaa aaaatactaa tgcttgggcc ccaataaagc 186840
aataaattca gaatctctgt gcatgtggcc ctaggcatca gtattgaaca catcactaag 186900
gtttgaaagc tcctccaggt gattctgatg cacccaagct taagcatcat tagttcaggg 186960
catggttata cgtgcacttt ggttcaatca gattgctatc ctaatcctag gttgaaaaca 187020
cctgcttggg agtaggaaat ttatttaggg gctggaggga ggtaagccgt ttagccctca 187080
agaagccttt aggggtgtgt ctaaggcaaa cctgagtggg taccatgagg ,gaagttaggg 187140
agatgtgaga gtacggggag aggcagagaa cacccctatc cacatgccat ccttctcctg 187200
gcaagctcct actattcctt tcagcacact cctagtcttg tccagccccc ctgagaagct 187260
ttccttcaac tgccactacc ctcagctcga tatcactttc tatgctcctg gctgcataat 187320
actttgtggg tatcttcaac attgcactca gcatgtatgt taccatgatc tcgtctcacc 187380
catcttagtg caaatggagg tttcatctta ttaattttgg ggtctctgga gcttagtgca 187440
gtgtttgaca cttagcagat atttaatgaa tgattatatg aatatgaaaa aaacgcatag 187500
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aatatgtaaa ttaggagtgt attaaggaca aatctgcgta ttagacaaat atttaagggt 187560
ttcttgtact tttagcattg taatgtgata gcttagtagg tattcagtgt tacaaaagtt 187620
cttttaggct aacctgatga cctagccatc atttttaata ttttttctct aagataaatg 187680
aaattcattg tgtatttcaa ataaccaact tccagatgaa tctttagatc acaatgtcct 187740
cctgatttaa ggactgccta tacttttcaa aatgccccat gctgagtctt gagtatccct 187800
cttgtaactt ctaccaatta tttaactgag ttgtccagaa aaagcttatt tccaaaaatt 187860
tattctagac aaaaagaacc acccccccca acacacacac acatacacac acacactttt 187920
accctttaac aggtaaattc tatgagctaa gtctggatct acaccatttc aggccttttt 187980
aaaatgtggc acttaggcca ggtgctgtgg ctcatgcctg taaccccaac actttgggag 188040
gccgaggtgg gagaatcact tgagcccagg ggttcaaaac cagcctggga aacatagtgg 188100
gtaccccatc tctacaaaat ttaaaaaatt agctgagctt gctggcacac atttgtggtc 188160
ctagctactc gagaggctga ggaggaggat cacttgggcc tggaaggtcg aggctgcagt 188220
gagctatgat tacaccactg cactccagcc tgggcaacag agcaaggccc tgcctcaaaa 188280
atgtagcact taattgttct tttctaataa tcttatcagt tggtggggta gatattcagc 188340
cagtgtttaa acaatggcta atagtcacat ggtgcctacc acatgccagg cactgcccta 188400
agtgccacca tttttcagtg tccttttctt cttcctagga gcctcaaaaa aaacccatat 188460
ctggccctgc actaaagaca gaaaaatcca caaagaggct ggatgtgagg acaaaattat 188520
cctcatttta tccagaatgg agccagtagt tttcttttcc ccaccaccaa ctaaaacata 188580
aaatgggtca gatgcagtgg ctcatgccgg taatcccagc actttgggag gccgaggcgg 188640
gcagatcatg aggtcaggag ttcgagacca acctggccaa catggtgaaa ccccgtctct 188700
actaaaaaca caaaaattag ctgggcatga tggtgcgcac ctgtaatccc agctacttgg 188760
gaggctgagg caagagaatc atttgaacct gggaggcgga ggttgcagtg agccgagatc 188820
gtgccactgc actccagcct gggcaataga gtgagactcc atctcaaaaa gaaagaaaga 188880
aaaaaaaaac agcatgaaac aatataaata aactagaacc caataaatcg tgaaggcccc 188940
agagagggag ccaccatcca gctgcagagg tggccacatc cccaccaggt gagtctaagt 189000
gctgctgcaa accagctcca tataccagca catgtgtctc agaccaagga agtttatgat 189060
gcagggagga gagccacctt aatggaatga gcaacaaggc agctgcagtc acagggtact 189120
tttgggggaa ataatttaca tttaaaaatt ttaaataagt agattgagta caaattgaga 189180
gcaaaggctt ctggctgaag agatgagaag caaagcatgg agcaatagaa tgacactaga 189240
tcatttcatt ctttcccttc agtgcttgtt gtttaagaga aggcatgaaa acatatttga 189300
tctgaaaaat aatgcattaa ctcattctac agctgagaag tcggcaagtc agggttctgc 189360
aagctctgag ctccccaccc tcaaaggact ttatgagcaa aggtgactcg tactttgggt 189420
gtctgcagaa gcaatacagt ttcctttaag taagaacaga gagtaaatga actaaaagcc 189480
tatttatttt ataaacagat aaccccaagg gagaatcact ttgtgaatgg tttttctaat 189540
acagctgtgt ctctgatgca ttgaaaaatc atgcccagct gggcgtggtg gctcacgcct 189600
gtaatcccag ccctttggga ggccgaagca ggcagattaa ttgggcccag cagttcaaga 189660
acaccctggg caacatggtg aaacctgcat ctctacaaaa aaaaaaacac aaaaaattgc 189720
ccgtgtgcgg cagtgcaagc ttttagtccc agctaccaaa aggctgaggt gggaaaatca 189780
tctgagccca ggaagtcgag gctgcagtga gccatgatcg cgccactgca ctccagcctg 189840
ggccacagag cgagactcca tctcaaaaaa aaagaaaaaa agaaaagaaa aattacacgc 189900
ttcagctgca aatatttgaa agaagaattt acttttccaa ggaaagggta gtcacgtttt 189960
cttgaatgcc gacactaatt tacggaggct gtgtgctttg catccacacc agattttcat 190020
ctaaagaact gcctttgctt ggtaaatgaa aatggagatt cttaagaagg atttgatagt 190080
ctttttcaaa accctcagat accagatgct cctaacgcat ctgcctttag tctgcccagg 190140
aatctcagga agtcatccat tcctcgagta cagtttcagt gatacaagga gatagcagcc 190200
cacattttat acgctattct gagttatacc agaggttcgt cgtttgggaa aatacggtta 190260
ttttaaccta gatgcaaatc ctatctgggt tactctgcag tttcagcatt gagagacttc 190320
ttcatgacag aaaagtggtt tcctttttct tatcagcaac atacactcag gccattttcc 190380
ccggaagaat ccagcttatc ttactactac aacatatatc tggtcattta tgacactaga 190440
atctagaaac taaaatgtta gtttaacaaa gtgaaaagcc aggcacggtg gctcatacct 190500
gtaatcccag cactttggga ggctgaggtg ggaggattgc ttgagcccag gagttcaaca 190560
ctagcctggg caacatagca atagcctcgt ctcgtcttcc tttttttttt ttttttttta 190620
agtgaaagaa gtcagtcttt aaaaagctac atactgtgag gtttcaactc tgtgacatcc 190680
tggaaaaggc agaactatgg aaacagtaaa aagattggtg attgttcagg gttgtgggga-190740
gagaggaatg aatagatgga gcatgggact tttagggcag taaagctatt ctacatgata 190800
ctgtaatgat ggatacatgt cattatacat gtgtcaaaac ccacagaatg tacaacacca 190860
atgtaatgta attgaacttt ggctaataac aacgtgtcaa tattggctca tcagttgtaa 190920
caaatgtacc ataccaaagc aagaagttac tagtggaaac tgagggtata agagaactct 190980
actttttgtt cacttttttt gttgtaaacc taaagctgct caaaaacatc tattcatatt 191040
tctaaaaaac agcaaaaaga cagtttagcc tcatggcata tccatggagg tgggggaggt 191100
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ggggacgggg tcattctagt ataattaagt cttctttggt acacaacata taggctcaag 191160
caatttcatt tgaaagtagt tatcagtata ttaaatctcc ttatgttttt agtggttgct 191220
gccattacta gtctcttgct ttaaaaaaag aagaggaaga atatgaaatt agcttaccct 191280
cttctcaaga agtttctttc agtcagagat tctgtcatac tcttcccaca ttccaaacag 191340
ttggcaactt tgcctcactt caggagcctg catactctac tttctaggtg tcttagccaa 191400
aaagaaaatc cgtgcttact gccctttatt aggtgtgatg taacccacaa aatcactggc 191460
tgtgcactaa ctcacagact ggatttgtgc agctcaggga ggagtgttta ttcctttccc 191520
atggtgcaga aaggaggatg gttggataat aaccacaata aatatctcgg ttatgaagag 191580
tccacacgtt atggaaacaa ctatcattag gtacagtaca agtagcgatg agtagttatg 191640
acttgtcaaa gaaaaaccca cacttacaaa tttacaaatc aaacaaagaa agatgtgtgc 191700
taaatgctgc attttgcccc ttataagatg catttccctc ttagggggaa aaaaaaaaca 191760
aggtatccaa aaattgccct gtattcgatg tgcctttaac tcagtgacag ccaaaaaaga 191820
gtaaactggc ataaagtcag aaggaaattc atactctctc atggtgtcag tggtgtatgc 191880
aggctcatga acaaaattaa gtttcagctg ggaatggtgg ctcacacctg taatcccagc 191940
actttgggag gccaaggcag gaggatcact tgagtccagg agtttgagac catcctgggc 192000
aatatgatga aaccccatct ctacaaaaaa atacaataat tagctgggca tggtggtgca 192060
cgcctgtagt cccaactgtt caggaggctg aggtgggagg atcacttgag cccagtagat 192120
cgaggctgca gtgagccaag attccaccac tgcactccag cctgggtgac atgagcgaga 192180
ccctatcatc tctgaaaaaa aaaaaaaaaa attaagtttt ctgcctagaa qatcacttgt 192240
tcatgtgtac taaagataat aatttttttc ttttttaaaa aatgtcttaa gaagtgggat 192300
gtgtcttact tatatacctg cgtatcttaa agagagggaa caagatcttc ctaggcccac 192360
aattgattct cttctgtcat attttcttgg acttgatttc ccaaatgtta cattggctca 192420
ctggtgtgct ccccagctca gtattcggtt tctgcctgct accaaaaata attggcaaga 192480
tagcagagta tttcacctcc atagtatctg aaagatgagg gatataacct caccctccct 192540
aattttctga gaaaattccg tgggctcttc ttatttaaga cctagcatgt taattaataa 192600
aagaaaatag aaagctgata agcaacttca gcaaagtctc aggatacaaa atcaacgtgc 192660
aaaaatcaca agcattcctt tacatcaaca atagacaagt gagagccaac tcatgagtga 192720
actcccattc acaattgcta caaagacagt aaaatacctg gaaatacaat ttacaagggc 192780
tgtgaaggaa ctcttcaagg agaactacaa accactgccc aaggaaataa gagaggatgt 192840
aaacaaatgg aaaaacattc catattcatg gataggaaga atcaatatag tgaaagtggc 192900
catactgccc aaagtaattt atagattcaa tgctattccc atccaactac cattgacatt 192960
cttcacagaa ttagaaaaag ctactttaaa tttcatatgg aaccaaggaa gaccccatat 193020
agccaagaca atcctaagca aaaagaacaa agctagaggc atcatgttac ctgacttcaa 193080
actatactac aaggctacag taaccaaaac agcatggtac tggtaccaaa acagacatat 193140
agaccaatgg cacagaacag agacctccga aataacacta cacatgtaca accatctgat 193200
cttcaacaaa cctcacagaa acaagagatg gggaaaagat ctcctattca aaaaaaggtg 193260
ctgtgaaaac tggctagcca tattcagaaa actgaaactg gaccccttcc ttacacttta 193320
tgcaaaatta actcaagatg gattaaagac ttaaatgtaa agcccaaaac cataaaaacc 193380
ctagaagaaa acctaggcaa taccactcag gacataggca tgggcaaata cttcatgatg 193440
aaaacaccaa aagcaatttc aacaaaagcc aaaattcaca aatgggatgt aattaaacta 193500
aagatcttca gcacagcaaa agaaactgtc atcagagtaa acaggcaacc tacagaatgg 193560
gagaacattt ttgcaatcta cccatctgac aaaggtctaa tatctagaat ttacaaggaa 193620
tgtaaacaaa tttaccagaa aaaaataaac aacttcatca acaagggggc aaaggatatg 193680
aacagatact tctcaaaaga agacatttac atggccaaca aacatatgaa gaaaagctca 193740
acatcactga tcatagagaa atgaaaatca aaactgcaat gagatactat ctcatgccag 193800
tcagaatggc tattattaaa aagtcaagaa acaatagatg ctagcgaggc tgtggagaaa 193860
taggaacact tttacactgt tggtgtgaat gtaaattagt tcaaccactg tggaagacag 193920
tacagcgatt cctcaaggat ctagaaccag aaataccatt tgacccagca atcccattac 193980
tgggtatgta cccaaaggaa tataaatcat tctactataa agacacatac acatgaatgt 194040
ttattgctgc actatttaca atagcaaaga cttggaacca acccaaatgc ctatcaatga 194100
tagactggat aaagcaaatg tggtacatgt acaccatgga atactatgca gccataaaaa 194160
aggaatgaga taatgtcctt tgcagggaca tggatgaagc tagaagccat tatcctcagc 194220
aaactaacac aggaacagaa aaccaaacac cgattgttct cactcataaa tgagagttga 194280
acaatgacaa cacatggaca tagggagggg aacaacacac accagggcct gtttagggga 194340
tggggagtga ggggagggaa cttagaggat gggtcaatag gtgcagcaaa ccaccatggc 194400
acatgtatac ctatgtaaca aacctgcacg ttctgcacat gtatctcgga actgaaagta 194460
aaatacaaaa taaataaaaa ataaaacact tagcatgaac ataattaatt aaagaaaata 194520
gggagtaaca aatctaagta aatatattgt tgcacttaac aacagttttt tttctcttta 194580
cttatctgat agagacaggg tcttgctatg ttgcccaggc tggtctcaaa ctcctaggtt 194640
caagaaatcc tccaacctca gcctcccaaa gtgctgggat tacaggtgta agacaccctg 194700
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ctcagcctgc catacatttt aactttggaa tgaaatttaa atttccattt aaacatagat 194760
gtttctgaaa ttatgggaat ttacaaaaca aaataacaag tcactcttgt aaagagatga 194820
gaaactgctc aatttttcta tatcaccctc tccacccttt gcctgggata agaggaggta 194880
acccagcttt tcccacactg agcagaggtg acctgcctct ctctaatgaa ctgcgcttta 194940
tgcctgtttt gttcctgaga gaagagtttg gatagaggag gattaaagtt tgttttaaga 195000
gacccttcct gcagggtata gtggctcatg cctgtaatcc cagcactttg ggaggccaag 195060
acaagaggat tgcttaagcc caggagtttg agaccagcca gggcaacgca ggaggacacc 195120
cccatctcta caaatattta aaaaaaaaaa aatagctgag catggcagta cacacttgta 195180
gtcccaagta gcaactcagg aggctgaggt aggaggatca cttgaacctg cgaagtggag 195240
gctgcagtga gttatcattg taccagtgta ctcacctggg caacatagca agaccctatc 195300
tctacaaaaa taaaaataaa aattagctgt gtatggtggt gtgcacctgt agtcctagct 195360
actcaggagg ctgaggtggg aagattgctt gagcccagga gtttgaggct gcagtgaact 195420
attactatgc cactgcactc cagcctgggc aaccgagcaa gacctaatct ctataaaaat 195480
gaaaaagacc cttccttcca acaacgttaa accactagag atgtttcgta ctccctacaa 195540
ttttttctct ctttctgttc acaatacatt gggacttaca ggcagagtaa gagtcaatgg 195600
aagctatggt aagaaagtca acagagggct ggaggaacaa taccaacagc taggattctc 195660
tactcccaga tttgctaaca agcattgatt tgctccactg atatacaaca caatctgagg 195720
gaagatcatt gaaaatctag gatcttttgg tctaaagttg gaatcatggg gaggcatttt 195780
ctcaaccttg ttcatgtctc tttcctttgc agggcaacag gctttggctt cttaaatgcg 195840
ctatgcaagg cagcagccgt cctgggaaac ttaatatttg gctctctggt cagcatcacc 195900
aaatcaatcc ccatcctgct ggcttctact gtgctcgtgt gtggaggact cgttgggctg 195960
tgcctgcctg acacacgaac ccaggttctg atgtaatggg aaaaaaagcc atccttcctg 196020
cgtttcttcc tcctgccctg ggtcaattct ccttcctgac tcaaggcttc agagttttcc 196080
tatatagaaa ggtgatcaag tatcagaaca taaacacgtg ctgtgactta aaatttagaa 196140
gcatatcatc ttgccccttt gtgattttgc acaggttgtt tgtttgtttt gttttgtttg 196200
aatgcattgt tatctttcca aactgtgatg ctactgcctc ccgcaaatca gtggaagcat 196260
ttctaaaatg ccctaggcag ccaggcttcc ctggggttct gctcattgaa agtttagaag 196320
ctaagaagac tgggctggtg taagaaatag atcctgctcc taatttaaca ttaacaggaa 196380
atataagtcg gcacattatt gcatttgtgc tgagaagagg gattcttttt tttttttttt 196440
ctaacagagt gatatttcct gtttacttag aaaaggacac ccagtaattc tcactgttat 196500
agcagagcct ttcaaagaaa accatgtggc accaaacagg gctgggtggg gctctttggg 196560
gcaagacaat taaggtgacc gctgacaaga aaaataaaag ctctgcaatt agcagcaaaa 196620
atctgagtgt gctgcataag caagatttct gtcagaaagc ccaattaaac ctctgaaaag 196680
tgtgaggaaa aagaactgta acagtatctg tgactgtggc atgtggttca gaatgtttga 196740
aaatcagaga tccgagagag agcttctgcc tccatttacc actcccactc cttcagctgc 196800
ccttggtctt gccagacagc agattcaggg aaagaaagct gcttcttaaa aactgtctgc 196860
ctacctgatt ctgccactta cccacactgt ctctagtcta gagattgcca gtggatagct 196920
aaattgtaaa ttgcaaatta ggaagaaatt cacacccaga gccatttgga aatacaaaat 196980
ggtacctctt tccagtacct ctttgctgtg ggactggccc gccaggaccc ccaccccacc 197040
tggcccgcag ttgacattca gcttcccagc tggcatcagc aaagagggtt tgtccctcta 197100
gcttccccag ggctggccca ctgctcaatg cgctcctggc tccctgtaaa atcacagccc 197160
gcctgcactc ctgagggccc ctctcacaga cagaccatct ccctgtggtt gtttgactgc 197220
tcattttacc tctggaatct gcctgggctt ggaagaaaga ggtcagccag gacattccca 197280
cgggcccgct gtcagctaca tgaccttctc gcctctcagg ctccccttcc cccaggctcg 197340
tcctttttac acctcttctt tgaaatctga accttccttg tagacttgca gccaatgacc 197400
agaagccagg aataaatccc atatttaaac aatattcctt ttttaactgc ctcgatagag 197460
gtttattctt gttacttatt ataggcattt tcaggcaagc catagcttgg ggcctccagg 197520
aagggaaagc ttttgtttcc actgatcatt attttcagag atgtcctaac tccccctgag 197580
gcccccctgc ctccccctac ccttcctaat gatccctcgc agagctcaga ctaagtgaaa 197640
agcttctttc attaacataa gccattaatc gacttcccct tgaagtaatt caggtctgca 197700
gaggggggaa gactggtgtt gggggcagtg ggaagagggt ggttccctgg gggtctgtga 197760
tcagggctag aacatcagga aaacccgaac caaaaactgc aaaggtggca ggaagaagaa 197820
aaatcactga aggctagaca agatgcattt gaaagatacc aaccgcacag ggaaagaaac 197880
aactttctct cacttctgcc tcctatcatt ttattggaga acaaagacag cagtcaaaca 197940
acccaagaaa gtaccctgta tgtttttctt cttccacatg tacccatttt cccacct 197997
<210> 4
<2l1> 727
<212> PRT
<213> Rattus norvegicus
5g
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<400> 4
Met Glu Asp Ser Tyr Lys Asp Arg Thr Ser Leu Met Lys G1y Ala Lys
1 5 10 15
Asp Ile Ala Lys Glu Val Lys Lys Gln Thr Val Lys Lys Val Asn Gln
20 25 30
Ala Val Asp Arg Ala Gln Asp Glu Tyr Thr Gln Arg Ser Tyr Ser Arg
35 40 45
Phe Gln Asp Glu Asp Asp Asp Asp Asp Tyr Tyr Pro Pro Gly Glu Thr
50 55 60
Tyr Ser Gly Glu Ala Asn Asp Asp Glu Gly Ser Ser Glu Ala Thr Glu
65 70 75 80
Gly His Asp Glu Glu Asp Glu Ile Tyr Glu Gly Glu Tyr Gln Gly Ile
0
85 90 95
Pro Ser Thr Asn Gln Gly Lys Asp Ser Ile Val Ser Val Gly Gln Pro
100 105 110
Lys Gly Asp Glu Tyr Lys Asp Arg Arg Glu Leu Glu Ser Glu Arg Arg
115 120 125
Ala Asp Glu Glu Glu Leu Ala Gln Gln Tyr Glu Leu Ile Ile Gln Glu
130 135 140
Cys Gly His Gly Arg Phe Gln Trp Ala Leu Phe Phe Val Leu G1y Met
145 150 155 160
Ala Leu Met Ala Asp Gly Val Glu Val Phe Val Val Gly Phe Val Leu
165 170 175
Pro Ser Ala Glu Thr Asp Leu Cys Ile Pro Asn Ser Gly Ser Gly Trp
180 185 190
Leu Gly Ser Ile Val Tyr Leu Gly Met Met Val Gly Ala Phe Phe Trp
195 200 205
Gly Gly Leu Ala Asp Lys Val Gly Arg Lys Gln Ser Leu Leu Ile Cys
210 215 220
Met Ser Val Asn Gly Phe Phe Ala Phe Leu Ser Ser Phe Val Gln Gly
225 230 235 240
Tyr Gly Phe Phe Leu Leu Cys Arg Leu Leu Ser Gly Phe Gly Ile Gly
245 250 255
Gly Ala Ile Pro Thr Val Phe Sex Tyr Phe Ala Glu Val Leu Ala Arg
260 265 270
Glu Lys Arg Gly Glu His Leu Ser Trp Leu Cys Met Phe Trp Met Ile
275 280 285
Gly Gly Ile Tyr Ala Ser Ala Met Ala Trp Ala Ile Ile Pro His Tyr
290 295 300
Gly Trp Ser Phe Ser Met Gly Ser Ala Tyr Gln Phe His Ser Trp Arg
305 310 315 320
Val Phe Val Ile Val Cys A1a Leu Pro Cys Val Ser Ser Val Val Ala
325 330 335
Leu Thr Phe Met Pro Glu Ser Pro Arg Phe Leu Leu Glu Val Gly Lys
340 345 350
His Asp Glu Ala Trp Met Tle Leu Lys Leu Ile His Asp Thr Asn Met
355 360 365
Arg Ala Arg Gly Gln Pro Glu Lys Val Phe Thr Val Asn Lys Ile Lys
370 375 380
Thr Pro Lys Gln Ile Asp Glu Leu Ile Glu Ile Glu Ser Asp Thr Gly
385 390 395 400
Thr Trp Tyr Arg Arg Cys Phe Val Arg Tle Arg Thr Glu Leu Tyr Gly
405 410 415 .
Ile Trp Leu Thr Phe Met Arg Cys Phe Asn Tyr Pro Val Arg Glu Asn
420 425 430
Thr Ile Lys Leu Thr Ile Val Trp Phe Thr Leu Ser Phe G1y Tyr Tyr
435 440 445
Gly Leu Ser Val Trp Phe Pro Asp Val Tle Lys His Leu G1n Ser Asp
450 455 460
59
CA 02439401 2003-08-25
WO 02/083900 PCT/US02/05520
Glu Tyr Ala Leu Leu Thr Arg Asn Val Gln Lys Asp Lys Tyr Ala Asn
465 470 475 480
Phe Ser Tle Asn Phe Thr Met Glu Asn Gln Val His Thr Gly Met Glu
485 490 495
Tyr Asp Asn Gly Arg Phe Leu Gly Val Lys Phe Lys Ser Val Thr Phe
500 505 510
Lys Asp Ser Val Phe Lys Ser Cys Thr Phe Asp Asp Val Thr Ser Val
515 520 525
Asn Thr Tyr Phe Lys Asn Cys Thr Phe Ile Asp Thr Leu Phe Glu Asn
530 535 540
Thr Asp Phe Glu Pro Tyr Lys Phe Ile Asp Ser Glu Phe Gln Asn Cys
545 550 555 560
Ser Phe Leu His Asn Lys Thr Gly Cys Gln Ile Thr Phe Asp Asp Asp
565 570 575
Tyr Ser Ala Tyr Trp Ile Tyr Phe Val Asn Phe Leu Gly Thr Leu Ala
580 585 590
Val Leu Pro Gly Asn Ile Val Ser Ala Leu Leu Met Asp Arg Ile Gly
595 600 605
Arg Leu Thr Met Leu Gly Gly Ser Met Val Leu Ser Gly Ile Ser Cys
6l0 6l5 620
Phe Phe Leu Trp Phe Gly Thr Ser Glu Ser Met Met Ile Gly Met Leu
625 630 635 640
Cys Leu Tyr Asn Gly Leu Thr Ile Ser Ala Trp Asn Ser Leu Asp Val
645 650 655
Val Thr Val Glu Leu Tyr Pro Thr Asp Arg Arg Ala Thr Gly Phe Gly
660 665 670
Phe Leu Asn Ala Leu Cys Lys Ala Ala Ala Val Leu Gly Asn Leu Ile
675 680 685
Phe Gly Ser Leu Val Ser Ile Thr Lys Ala Ile Pro Ile Leu Leu Ala
690 695 700
Ser Thr Val Leu Val Cys Gly Gly Leu Val Gly Leu Arg Leu Pro Asp
705 710 715 720
Thr Arg Thr Gln Val Leu Met
725
